Compositions and methods for treating acute radiation syndrome

ABSTRACT

Embodiments are directed to a method of treating acute radiation syndrome comprising administering to a subject following exposure to radiation a PIF peptide. Some embodiments describe a method of treating acute radiation syndrome following radiation exposure comprising transplanting bone marrow that has been exposed to a PIF peptide prior to transplantation into a subject. Other embodiments describe a method of increasing engraftment of a transplanted organ, tissue, or cell by pre-exposing the organ, tissue, or cell to a PIF peptide.

CROSS-REFERENCE TO RELATED APPLICATIONS

This application claims priority to and benefit of U.S. ProvisionalApplication Ser. No. 62/051,077 filed Sep. 16, 2014, entitled “Methodsfor Treating Acute Radiation Syndrome,” U.S. Provisional ApplicationSer. No. 62/113,298 filed Feb. 6, 2015, entitled “PIF Binding as aMarker for Immune Dysregulation,” and U.S. Provisional Application Ser.No. 62/211,660 filed Aug. 28, 2015, entitled “Compositions and Methodsfor the Treatment of Neurodamage,”. The contents of each applicationwhich are incorporated herein by reference in their respectiveentireties.

SUMMARY

In an embodiment, a method of treating acute radiation syndrome in asubject in need thereof after the subject has been exposed to radiationmay comprise administering a therapeutically effective amount of aPreImplantation Factor (PIF) peptide selected from SEQ ID NO: 1, SEQ IDNO: 2, SEQ ID NO: 3, SEQ ID NO: 4, SEQ ID NO: 5, SEQ ID NO: 6, SEQ IDNO: 7, SEQ ID NO: 8, SEQ ID NO: 9, SEQ ID NO: 10, SEQ ID NO: 11, SEQ IDNO: 12, SEQ ID NO: 13, SEQ ID NO: 14, SEQ ID NO: 15, SEQ ID NO: 16, SEQID NO: 17, SEQ ID NO: 18, SEQ ID NO: 19, SEQ ID NO: 20, SEQ ID NO: 21,SEQ ID NO: 22, SEQ ID NO: 23, SEQ ID NO: 24, SEQ ID NO: 25, SEQ ID NO:26, SEQ ID NO: 27, SEQ ID NO: 28, SEQ ID NO: 29, mimetics thereof, andcombinations thereof.

In an embodiment, a pharmaceutical composition comprising atherapeutically effective amount of a PIF peptide selected from SEQ IDNO: 1, SEQ ID NO: 2, SEQ ID NO: 3, SEQ ID NO: 4, SEQ ID NO: 5, SEQ IDNO: 6, SEQ ID NO: 7, SEQ ID NO: 8, SEQ ID NO: 9, SEQ ID NO: 10, SEQ IDNO: 11, SEQ ID NO: 12, SEQ ID NO: 13, SEQ ID NO: 14, SEQ ID NO: 15, SEQID NO: 16, SEQ ID NO: 17, SEQ ID NO: 18, SEQ ID NO: 19, SEQ ID NO: 20,SEQ ID NO: 21, SEQ ID NO: 22, SEQ ID NO: 23, SEQ ID NO: 24, SEQ ID NO:25, SEQ ID NO: 26, SEQ ID NO: 27, SEQ ID NO: 28, SEQ ID NO: 29, mimeticsthereof, and combinations thereof, may be used for the treatment ofacute radiation syndrome.

In an embodiment, a pharmaceutical composition comprising atherapeutically effective amount of a PIF peptide selected from SEQ IDNO: 1, SEQ ID NO: 2, SEQ ID NO: 3, SEQ ID NO: 4, SEQ ID NO: 5, SEQ IDNO: 6, SEQ ID NO: 7, SEQ ID NO: 8, SEQ ID NO: 9, SEQ ID NO: 10, SEQ IDNO: 11, SEQ ID NO: 12, SEQ ID NO: 13, SEQ ID NO: 14, SEQ ID NO: 15, SEQID NO: 16, SEQ ID NO: 17, SEQ ID NO: 18, SEQ ID NO: 19, SEQ ID NO: 20,SEQ ID NO: 21, SEQ ID NO: 22, SEQ ID NO: 23, SEQ ID NO: 24, SEQ ID NO:25, SEQ ID NO: 26, SEQ ID NO: 27, SEQ ID NO: 28, SEQ ID NO: 29, mimeticsthereof, and combinations thereof, may be used for the manufacture of amedicament for treating acute radiation syndrome.

In an embodiment, a method of treating acute radiation syndromefollowing radiation exposure may comprise transplanting one or aplurality of bone marrow cells into a subject in need thereof, whereinthe bone marrow is pre-exposed to a therapeutically effective amount ofPIF peptide prior to transplantation, and wherein the PIF peptideselected from SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, SEQ ID NO: 4,SEQ ID NO: 5, SEQ ID NO: 6, SEQ ID NO: 7, SEQ ID NO: 8, SEQ ID NO: 9,SEQ ID NO: 10, SEQ ID NO: 11, SEQ ID NO: 12, SEQ ID NO: 13, SEQ ID NO:14, SEQ ID NO: 15, SEQ ID NO: 16, SEQ ID NO: 17, SEQ ID NO: 18, SEQ IDNO: 19, SEQ ID NO: 20, SEQ ID NO: 21, SEQ ID NO: 22, SEQ ID NO: 23, SEQID NO: 24, SEQ ID NO: 25, SEQ ID NO: 26, SEQ ID NO: 27, SEQ ID NO: 28,SEQ ID NO: 29, mimetics thereof, and combinations thereof.

In an embodiment, a method of treating and/or preventing a heartdisorder or heart failure may comprise transplanting one or a pluralityof heart cells into a subject in need thereof, wherein the heart cellsare pre-exposed to a therapeutically effective amount of a PIF peptideprior to transplantation, and wherein the PIF peptide selected from oneor a combination of SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, SEQ ID NO:4, SEQ ID NO: 5, SEQ ID NO: 6, SEQ ID NO: 7, SEQ ID NO: 8, SEQ ID NO: 9,SEQ ID NO: 10, SEQ ID NO: 11, SEQ ID NO: 12, SEQ ID NO: 13, SEQ ID NO:14, SEQ ID NO: 15, SEQ ID NO: 16, SEQ ID NO: 17, SEQ ID NO: 18, SEQ IDNO: 19, SEQ ID NO: 20, SEQ ID NO: 21, SEQ ID NO: 22, SEQ ID NO: 23, SEQID NO: 24, SEQ ID NO: 25, SEQ ID NO: 26, SEQ ID NO: 27, SEQ ID NO: 28,SEQ ID NO: 29, mimetics thereof, and combinations thereof.

In an embodiment, a method of treating and/or preventing an adrenal celldisorder may comprise transplanting one or a plurality of adrenal cellsinto a subject in need thereof, wherein the adrenal cells arepre-exposed to a therapeutically effective amount of a PIF peptide priorto transplantation, and wherein the PIF peptide selected from one or acombination of SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, SEQ ID NO: 4,SEQ ID NO: 5, SEQ ID NO: 6, SEQ ID NO: 7, SEQ ID NO: 8, SEQ ID NO: 9,SEQ ID NO: 10, SEQ ID NO: 11, SEQ ID NO: 12, SEQ ID NO: 13, SEQ ID NO:14, SEQ ID NO: 15, SEQ ID NO: 16, SEQ ID NO: 17, SEQ ID NO: 18, SEQ IDNO: 19, SEQ ID NO: 20, SEQ ID NO: 21, SEQ ID NO: 22, SEQ ID NO: 23, SEQID NO: 24, SEQ ID NO: 25, SEQ ID NO: 26, SEQ ID NO: 27, SEQ ID NO: 28,SEQ ID NO: 29, mimetics thereof, and combinations thereof.

In an embodiment, a method of treating and/or preventing a blooddisorder may comprise transplanting one or a plurality of hematopoieticcells into a subject in need thereof, wherein the hematopoietic cellsare pre-exposed to a therapeutically effective amount of a PIF peptideprior to transplantation, and wherein the PIF peptide selected from oneor a combination of SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, SEQ ID NO:4, SEQ ID NO: 5, SEQ ID NO: 6, SEQ ID NO: 7, SEQ ID NO: 8, SEQ ID NO: 9,SEQ ID NO: 10, SEQ ID NO: 11, SEQ ID NO: 12, SEQ ID NO: 13, SEQ ID NO:14, SEQ ID NO: 15, SEQ ID NO: 16, SEQ ID NO: 17, SEQ ID NO: 18, SEQ IDNO: 19, SEQ ID NO: 20, SEQ ID NO: 21, SEQ ID NO: 22, SEQ ID NO: 23, SEQID NO: 24, SEQ ID NO: 25, SEQ ID NO: 26, SEQ ID NO: 27, SEQ ID NO: 28,SEQ ID NO: 29, mimetics thereof, and combinations thereof.

In an embodiment, a method of increasing the viability of an organ,tissue, or cell prior to its transplantation into a subject in need oftransplantation may comprise treating the organ, tissue or cell with atherapeutically effective amount of PIF peptide prior totransplantation, wherein the PIF peptide is selected from one or acombination of SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, SEQ ID NO: 4,SEQ ID NO: 5, SEQ ID NO: 6, SEQ ID NO: 7, SEQ ID NO: 8, SEQ ID NO: 9,SEQ ID NO: 10, SEQ ID NO: 11, SEQ ID NO: 12, SEQ ID NO: 13, SEQ ID NO:14, SEQ ID NO: 15, SEQ ID NO: 16, SEQ ID NO: 17, SEQ ID NO: 18, SEQ IDNO: 19, SEQ ID NO: 20, SEQ ID NO: 21, SEQ ID NO: 22, SEQ ID NO: 23, SEQID NO: 24, SEQ ID NO: 25, SEQ ID NO: 26, SEQ ID NO: 27, SEQ ID NO: 28,SEQ ID NO: 29, mimetics thereof, and combinations thereof.

In an embodiment, a method of increasing the likelihood of acceptance ofa transplant of a donor organ, tissue, or cell into a subject maycomprise exposing the organ, tissue or cell to one or more compositionscomprising at least one PIF peptide or a mutant thereof or apharmaceutically acceptable salt thereof prior to transplanting theorgan, tissue, or cell into the subject, wherein the PIF peptide isselected from one or a combination of SEQ ID NO: 1, SEQ ID NO: 2, SEQ IDNO: 3, SEQ ID NO: 4, SEQ ID NO: 5, SEQ ID NO: 6, SEQ ID NO: 7, SEQ IDNO: 8, SEQ ID NO: 9, SEQ ID NO: 10, SEQ ID NO: 11, SEQ ID NO: 12, SEQ IDNO: 13, SEQ ID NO: 14, SEQ ID NO: 15, SEQ ID NO: 16, SEQ ID NO: 17, SEQID NO: 18, SEQ ID NO: 19, SEQ ID NO: 20, SEQ ID NO: 21, SEQ ID NO: 22,SEQ ID NO: 23, SEQ ID NO: 24, SEQ ID NO: 25, SEQ ID NO: 26, SEQ ID NO:27, SEQ ID NO: 28, SEQ ID NO: 29, mimetics thereof, and combinationsthereof.

A method of reducing the likelihood of rejection of an engrafted tissuemay comprise exposing the engrafted tissue to one or more pharmaceuticalcompositions comprising a therapeutically effective amount of at leastone PIF peptide or a mutant thereof or a pharmaceutically acceptablesalt thereof prior to transplanting the tissue into a subject, whereinthe PIF peptide is selected from one or a combination of SEQ ID NO: 1,SEQ ID NO: 2, SEQ ID NO: 3, SEQ ID NO: 4, SEQ ID NO: 5, SEQ ID NO: 6,SEQ ID NO: 7, SEQ ID NO: 8, SEQ ID NO: 9, SEQ ID NO: 10, SEQ ID NO: 11,SEQ ID NO: 12, SEQ ID NO: 13, SEQ ID NO: 14, SEQ ID NO: 15, SEQ ID NO:16, SEQ ID NO: 17, SEQ ID NO: 18, SEQ ID NO: 19, SEQ ID NO: 20, SEQ IDNO: 21, SEQ ID NO: 22, SEQ ID NO: 23, SEQ ID NO: 24, SEQ ID NO: 25, SEQID NO: 26, SEQ ID NO: 27, SEQ ID NO: 28, SEQ ID NO: 29, mimeticsthereof, and combinations thereof.

A method of increasing production of hematopoietic cells in a subjectmay comprise administering one or more pharmaceutical compositionscomprising a therapeutically effective amount of at least one PIFpeptide or a mutant thereof or a pharmaceutically acceptable saltthereof, wherein the PIF peptide is selected from one or a combinationof SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, SEQ ID NO: 4, SEQ ID NO: 5,SEQ ID NO: 6, SEQ ID NO: 7, SEQ ID NO: 8, SEQ ID NO: 9, SEQ ID NO: 10,SEQ ID NO: 11, SEQ ID NO: 12, SEQ ID NO: 13, SEQ ID NO: 14, SEQ ID NO:15, SEQ ID NO: 16, SEQ ID NO: 17, SEQ ID NO: 18, SEQ ID NO: 19, SEQ IDNO: 20, SEQ ID NO: 21, SEQ ID NO: 22, SEQ ID NO: 23, SEQ ID NO: 24, SEQID NO: 25, SEQ ID NO: 26, SEQ ID NO: 27, SEQ ID NO: 28, SEQ ID NO: 29,mimetics thereof, and combinations thereof.

A method of increasing the likelihood of successful engraftment of atransplanted organ, tissue, or cells may comprise transplanting anorgan, tissue, or cell into a subject in need thereof, wherein theorgan, tissue, or cell is pre-exposed to a PIF peptide prior totransplantation, wherein the PIF peptide is selected from one or acombination of SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, SEQ ID NO: 4,SEQ ID NO: 5, SEQ ID NO: 6, SEQ ID NO: 7, SEQ ID NO: 8, SEQ ID NO: 9,SEQ ID NO: 10, SEQ ID NO: 11, SEQ ID NO: 12, SEQ ID NO: 13, SEQ ID NO:14, SEQ ID NO: 15, SEQ ID NO: 16, SEQ ID NO: 17, SEQ ID NO: 18, SEQ IDNO: 19, SEQ ID NO: 20, SEQ ID NO: 21, SEQ ID NO: 22, SEQ ID NO: 23, SEQID NO: 24, SEQ ID NO: 25, SEQ ID NO: 26, SEQ ID NO: 27, SEQ ID NO: 28,SEQ ID NO: 29, mimetics thereof, and combinations thereof.

A method of treating and/or preventing adult or juvenile type I or typeII diabetes may comprise transplanting one or a plurality of pancreaticislet cells into a subject in need thereof, wherein the islet cells arepre-exposed to a therapeutically effective amount of PIF peptide priorto transplantation, and wherein the PIF peptide is selected from one ora combination of SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, SEQ ID NO: 4,SEQ ID NO: 5, SEQ ID NO: 6, SEQ ID NO: 7, SEQ ID NO: 8, SEQ ID NO: 9,SEQ ID NO: 10, SEQ ID NO: 11, SEQ ID NO: 12, SEQ ID NO: 13, SEQ ID NO:14, SEQ ID NO: 15, SEQ ID NO: 16, SEQ ID NO: 17, SEQ ID NO: 18, SEQ IDNO: 19, SEQ ID NO: 20, SEQ ID NO: 21, SEQ ID NO: 22, SEQ ID NO: 23, SEQID NO: 24, SEQ ID NO: 25, SEQ ID NO: 26, SEQ ID NO: 27, SEQ ID NO: 28,SEQ ID NO: 29, mimetics thereof, and combinations thereof.

In any of the above-described embodiments, the subject may or may notreceive a bone marrow transplant (BMT). In any of the above-describedembodiments, the acute radiation syndrome may be caused by exposure tolethal or sub-lethal radiation. In any of the above-describedembodiments, the acute radiation syndrome may be caused by exposure to aradiation dose of from about 100 rads to about 6000 rads. In any of theabove-described embodiments, the acute radiation syndrome may or may notcomprise delayed effects of acute radiation exposure, including damageto any organ, tissue, or cell.

In an embodiment, a method of increasing the likelihood of a transplantrecipient's acceptance of donor tissue may comprise exposing the donortissue to one or more compositions comprising a PIF peptide or a mutantthereof prior to transplanting the tissue into the recipient.

In an embodiment, a method of reducing the likelihood of rejection ofengrafted tissue may comprise exposing the tissue to one or morepharmaceutical compositions comprising a therapeutically effectiveamount of a PIF peptide or a mutant thereof prior to transplanting thetissue.

In an embodiment, a method of increasing the production of hematopoieticcells in a subject with a depleted number of hematopoietic cells maycomprise administering one or more pharmaceutical compositionscomprising a therapeutically effective amount of a PIF peptide or amutant thereof. In some embodiments, the hematopoietic cells may be redblood cells. In some embodiments, the hematopoietic cells may beplatelets.

In some embodiments, the step of administering to the subject at leastone PIF peptide, an analog thereof, or a pharmaceutically acceptablesalt thereof comprises administering a therapeutically effective dose ofthe at least one PIF molecule, an analog thereof, or a pharmaceuticallyacceptable salt thereof.

In some embodiments, the step of administering to the subject at leastone PIF peptide, an analog thereof, or a pharmaceutically acceptablesalt thereof comprises administering a therapeutically effective dose ofthe PIF peptide, an analog thereof, or pharmaceutically acceptable saltthereof from about 0.001 mg/kg to about 200 mg/kg.

In some embodiments, the step of administering to the subject at leastone PIF peptide, an analog thereof, or a pharmaceutically acceptablesalt thereof comprises administering a therapeutically effective dose ofthe PIF peptide, an analog thereof, or pharmaceutically acceptable saltthereof from about 0.5 mg/kg to about 5 mg/kg.

In some embodiments, the PIF peptide, analog thereof, orpharmaceutically acceptable salt thereof comprises a chemical targetingmoiety and/or a radioactive moiety.

In some embodiments, the at least one inhibitor of nuclear translocationof beta-catenin or pharmaceutically acceptable salt thereof comprises atleast one radioactive moiety comprising at least one or a combination ofthe following isotopes: ²H, ³H, ¹³C, ¹⁴C, ¹⁵N, ¹⁶O, ¹⁷O, ³¹P, ³²P, ³⁵S,¹⁸F, and ³⁶Cl.

In some embodiments, the method further comprises administering at leastone analgesic and/or one anti-inflammatory compound.

In some embodiments, the method further comprises administering at leastone analgesic and or one anti-inflammatory compound before, after, orsimultaneously with the administration of a therapeutically effectivedose of at least one PIF peptide, an analog thereof or pharmaceuticallyacceptable salt thereof.

In some embodiments, the therapeutically effective dose is from about1.0 mg/kg to about 5.5 mg/kg, wherein kg is kilograms of the subject andmg is milligrams of the therapeutically effective dose.

In some embodiments, the PIF peptide comprises SEQ ID NO: 1, SEQ ID NO:2, SEQ ID NO: 3, SEQ ID NO: 4, SEQ ID NO: 5, SEQ ID NO: 6, SEQ ID NO: 7,SEQ ID NO: 8, SEQ ID NO: 9, SEQ ID NO: 10, SEQ ID NO: 11, SEQ ID NO: 12,SEQ ID NO: 13, SEQ ID NO: 14, SEQ ID NO: 15, SEQ ID NO: 16, SEQ ID NO:17, SEQ ID NO: 18, SEQ ID NO: 19, SEQ ID NO: 20, SEQ ID NO: 21, SEQ IDNO: 22, SEQ ID NO: 23, SEQ ID NO: 24, SEQ ID NO: 25, SEQ ID NO: 26, SEQID NO: 27, SEQ ID NO: 28, SEQ ID NO: 29, mimetics thereof, orpharmaceutically acceptable salts thereof, and/or combinations thereof.In some embodiments, the PIF peptide comprises SEQ ID NO: 1, mimeticsthereof, or pharmaceutically acceptable salts thereof, and/orcombinations thereof. In some embodiments, the PIF peptide comprises SEQID NO: 2, mimetics thereof, or pharmaceutically acceptable saltsthereof, and/or combinations thereof. In some embodiments, the PIFpeptide comprises SEQ ID NO: 3, mimetics thereof, or pharmaceuticallyacceptable salts thereof, and/or combinations thereof. In someembodiments, the PIF peptide comprises SEQ ID NO: 4, mimetics thereof,or pharmaceutically acceptable salts thereof, and/or combinationsthereof. In some embodiments, the PIF peptide comprises SEQ ID NO: 5,mimetics thereof, or pharmaceutically acceptable salts thereof, and/orcombinations thereof. In some embodiments, the PIF peptide comprises SEQID NO: 6, mimetics thereof, or pharmaceutically acceptable saltsthereof, and/or combinations thereof. In some embodiments, the PIFpeptide comprises SEQ ID NO: 7, mimetics thereof, or pharmaceuticallyacceptable salts thereof, and/or combinations thereof. In someembodiments, the PIF peptide comprises SEQ ID NO: 8, mimetics thereof,or pharmaceutically acceptable salts thereof, and/or combinationsthereof. In some embodiments, the PIF peptide comprises SEQ ID NO: 9,mimetics thereof, or pharmaceutically acceptable salts thereof, and/orcombinations thereof. In some embodiments, the PIF peptide comprises SEQID NO: 10, mimetics thereof, or pharmaceutically acceptable saltsthereof, and/or combinations thereof. In some embodiments, the PIFpeptide comprises SEQ ID NO: 11, mimetics thereof, or pharmaceuticallyacceptable salts thereof, and/or combinations thereof. In someembodiments, the PIF peptide comprises SEQ ID NO: 12, mimetics thereof,or pharmaceutically acceptable salts thereof, and/or combinationsthereof. In some embodiments, the PIF peptide comprises SEQ ID NO: 13,mimetics thereof, or pharmaceutically acceptable salts thereof, and/orcombinations thereof. In some embodiments, the PIF peptide comprises SEQID NO: 14, mimetics thereof, or pharmaceutically acceptable saltsthereof, and/or combinations thereof. In some embodiments, the PIFpeptide comprises SEQ ID NO: 15, mimetics thereof, or pharmaceuticallyacceptable salts thereof, and/or combinations thereof. In someembodiments, the PIF peptide comprises SEQ ID NO: 16, mimetics thereof,or pharmaceutically acceptable salts thereof, and/or combinationsthereof. In some embodiments, the PIF peptide comprises SEQ ID NO: 17,mimetics thereof, or pharmaceutically acceptable salts thereof, and/orcombinations thereof. In some embodiments, the PIF peptide comprises SEQID NO: 18, mimetics thereof, or pharmaceutically acceptable saltsthereof, and/or combinations thereof. In some embodiments, the PIFpeptide comprises SEQ ID NO: 19, mimetics thereof, or pharmaceuticallyacceptable salts thereof, and/or combinations thereof. In someembodiments, the PIF peptide comprises SEQ ID NO: 20, mimetics thereof,or pharmaceutically acceptable salts thereof, and/or combinationsthereof. In some embodiments, the PIF peptide comprises SEQ ID NO: 21,mimetics thereof, or pharmaceutically acceptable salts thereof, and/orcombinations thereof. In some embodiments, the PIF peptide comprises SEQID NO: 21, mimetics thereof, or pharmaceutically acceptable saltsthereof, and/or combinations thereof. In some embodiments, the PIFpeptide comprises SEQ ID NO: 22, mimetics thereof, or pharmaceuticallyacceptable salts thereof, and/or combinations thereof. In someembodiments, the PIF peptide comprises SEQ ID NO: 23, mimetics thereof,or pharmaceutically acceptable salts thereof, and/or combinationsthereof. In some embodiments, the PIF peptide comprises SEQ ID NO: 24,mimetics thereof, or pharmaceutically acceptable salts thereof, and/orcombinations thereof. In some embodiments, the PIF peptide comprises SEQID NO: 25, mimetics thereof, or pharmaceutically acceptable saltsthereof, and/or combinations thereof. In some embodiments, the PIFpeptide comprises SEQ ID NO: 26, mimetics thereof, or pharmaceuticallyacceptable salts thereof, and/or combinations thereof. In someembodiments, the PIF peptide comprises SEQ ID NO: 27, mimetics thereof,or pharmaceutically acceptable salts thereof, and/or combinationsthereof. In some embodiments, the PIF peptide comprises SEQ ID NO: 28,mimetics thereof, or pharmaceutically acceptable salts thereof, and/orcombinations thereof. In some embodiments, the PIF peptide comprises SEQID NO: 29, mimetics thereof, or pharmaceutically acceptable saltsthereof, and/or combinations thereof.

The present disclosure also relates to a method of treating orpreventing acute radiation syndrome in a subject in need thereof, themethod comprising administering to the subject at least onepharmaceutical composition comprising: pre-implantation factor (PIF)peptide, an analog thereof, or a pharmaceutically acceptable saltthereof; and a pharmaceutically acceptable carrier.

In some embodiments, the pharmaceutically acceptable carrier is sterileand pyrogen-free water.

In some embodiments, the therapeutically effective dose of one or acombination of PIF peptide or analogs thereof or pharmaceuticallyacceptable salts thereof is about 0.2 mg/kg, wherein kg is kilograms ofthe subject and mg is milligrams of the therapeutically effective dose.In some embodiments, the therapeutically effective dose of one or acombination of PIF peptide or analogs thereof or pharmaceuticallyacceptable salts thereof is about 0.3 mg/kg, wherein kg is kilograms ofthe subject and mg is milligrams of the therapeutically effective dose.In some embodiments, the therapeutically effective dose of one or acombination of PIF peptide or analogs thereof or pharmaceuticallyacceptable salts thereof is about 0.4 mg/kg, wherein kg is kilograms ofthe subject and mg is milligrams of the therapeutically effective dose.In some embodiments, the therapeutically effective dose of one or acombination of PIF peptide or analogs thereof or pharmaceuticallyacceptable salts thereof is about 0.5 mg/kg, wherein kg is kilograms ofthe subject and mg is milligrams of the therapeutically effective dose.In some embodiments, the therapeutically effective dose of one or acombination of PIF peptide or analogs thereof or pharmaceuticallyacceptable salts thereof is about 0.6 mg/kg, wherein kg is kilograms ofthe subject and mg is milligrams of the therapeutically effective dose.In some embodiments, the therapeutically effective dose of one or acombination of PIF peptide or analogs thereof or pharmaceuticallyacceptable salts thereof is about 0.7 mg/kg, wherein kg is kilograms ofthe subject and mg is milligrams of the therapeutically effective dose.In some embodiments, the therapeutically effective dose of one or acombination of PIF peptide or analogs thereof or pharmaceuticallyacceptable salts thereof is about 0.75 mg/kg, wherein kg is kilograms ofthe subject and mg is milligrams of the therapeutically effective dose.In some embodiments, the therapeutically effective dose of one or acombination of PIF peptide or analogs thereof or pharmaceuticallyacceptable salts thereof is about 0.8 mg/kg, wherein kg is kilograms ofthe subject and mg is milligrams of the therapeutically effective dose.In some embodiments, the therapeutically effective dose of one or acombination of PIF peptide or analogs thereof or pharmaceuticallyacceptable salts thereof is about 0.9 mg/kg, wherein kg is kilograms ofthe subject and mg is milligrams of the therapeutically effective dose.In some embodiments, the therapeutically effective dose of one or acombination of PIF peptide or analogs thereof or pharmaceuticallyacceptable salts thereof is about 1.0 mg/kg, wherein kg is kilograms ofthe subject and mg is milligrams of the therapeutically effective dose.In some embodiments, the therapeutically effective dose of one or acombination of PIF peptide or analogs thereof or pharmaceuticallyacceptable salts thereof is about 1.5 mg/kg, wherein kg is kilograms ofthe subject and mg is milligrams of the therapeutically effective dose.In some embodiments, the therapeutically effective dose of one or acombination of PIF peptide or analogs thereof or pharmaceuticallyacceptable salts thereof is about 2.0 mg/kg, wherein kg is kilograms ofthe subject and mg is milligrams of the therapeutically effective dose.In some embodiments, the therapeutically effective dose of one or acombination of PIF peptide or analogs thereof or pharmaceuticallyacceptable salts thereof is about 3.0 mg/kg, wherein kg is kilograms ofthe subject and mg is milligrams of the therapeutically effective dose.In some embodiments, the therapeutically effective dose of one or acombination of PIF peptide or analogs thereof or pharmaceuticallyacceptable salts thereof is about 4.0 mg/kg, wherein kg is kilograms ofthe subject and mg is milligrams of the therapeutically effective dose.In some embodiments, the therapeutically effective dose of one or acombination of PIF peptide or analogs thereof or pharmaceuticallyacceptable salts thereof is about 5.0 mg/kg, wherein kg is kilograms ofthe subject and mg is milligrams of the therapeutically effective dose.In some embodiments, the therapeutically effective dose of one or acombination of PIF peptide or analogs thereof or pharmaceuticallyacceptable salts thereof is about 6.0 mg/kg, wherein kg is kilograms ofthe subject and mg is milligrams of the therapeutically effective dose.In some embodiments, the therapeutically effective dose of one or acombination of PIF peptide or analogs thereof or pharmaceuticallyacceptable salts thereof is about 7.0 mg/kg, wherein kg is kilograms ofthe subject and mg is milligrams of the therapeutically effective dose.In some embodiments, the therapeutically effective dose of one or acombination of PIF peptide or analogs thereof or pharmaceuticallyacceptable salts thereof is about 8.0 mg/kg, wherein kg is kilograms ofthe subject and mg is milligrams of the therapeutically effective dose.In some embodiments, the therapeutically effective dose of one or acombination of PIF peptide or analogs thereof or pharmaceuticallyacceptable salts thereof is about 9.0 mg/kg, wherein kg is kilograms ofthe subject and mg is milligrams of the therapeutically effective dose.In some embodiments, the therapeutically effective dose of one or acombination of PIF peptide or analogs thereof or pharmaceuticallyacceptable salts thereof is about 10.0 mg/kg, wherein kg is kilograms ofthe subject and mg is milligrams of the therapeutically effective dose.

In some embodiments, a PIF peptide, mimetics thereof, or combinationsthereof may be administered within about 24 hours after exposure toradiation. In some embodiments, a PIF peptide, mimetics thereof, orcombinations thereof may be administered intermittently or continuouslyfor about 2-14 days. In some embodiments, a PIF peptide, mimeticsthereof, or combinations thereof may be administered intermittently orcontinuously for about 2 days. In some embodiments, a PIF peptide,mimetics thereof, or combinations thereof may be administeredintermittently or continuously for about 3 days. In some embodiments, aPIF peptide, mimetics thereof, or combinations thereof may beadministered intermittently or continuously for about 4 days. In someembodiments, a PIF peptide, mimetics thereof, or combinations thereofmay be administered intermittently or continuously for about 5 days. Insome embodiments, a PIF peptide, mimetics thereof, or combinationsthereof may be administered intermittently or continuously for about 6days. In some embodiments, a PIF peptide, mimetics thereof, orcombinations thereof may be administered intermittently or continuouslyfor about 7 days. In some embodiments, a PIF peptide, mimetics thereof,or combinations thereof may be administered intermittently orcontinuously for about 8 days. In some embodiments, a PIF peptide,mimetics thereof, or combinations thereof may be administeredintermittently or continuously for about 9 days. In some embodiments, aPIF peptide, mimetics thereof, or combinations thereof may beadministered intermittently or continuously for about 10 days. In someembodiments, a PIF peptide, mimetics thereof, or combinations thereofmay be administered intermittently or continuously for about 11 days. Insome embodiments, a PIF peptide, mimetics thereof, or combinationsthereof may be administered intermittently or continuously for about 12days. In some embodiments, a PIF peptide, mimetics thereof, orcombinations thereof may be administered intermittently or continuouslyfor about 13 days. In some embodiments, a PIF peptide, mimetics thereof,or combinations thereof may be administered intermittently orcontinuously for about 14 days. In some embodiments, a PIF peptide,mimetics thereof, or combinations thereof may be administeredintermittently, the dosing regimen comprising about 1 dose per day orabout 1 dose every 2 days for at least about 12 weeks.

The present disclosure also relates to a pharmaceutical compositioncomprising (i) a therapeutically effective dose of one or a combinationof PIF peptide or analogs thereof or pharmaceutically acceptable saltsthereof; and (ii) a pharmaceutically acceptable carrier.

In some embodiments, the pharmaceutically acceptable carrier is sterileand pyrogen-free water or Lactated Ringer's solution.

In some embodiments, the composition further comprises a therapeuticallyeffective dose of one or a plurality of active agents.

In some embodiments, the one or plurality of active agents is one or acombination of compounds chosen from: an anti-inflammatory compound,alpha-adrenergic agonist, antiarrhythmic compound, analgesic compound,and an aesthetic compound.

In some embodiments, the composition further comprises one or aplurality of stem cells.

In some embodiments, the stem cell is an autologous stem cell.

In some embodiments, the pharmaceutical composition is administered viaparenteral injection, subcutaneous injection, intravenous injection,intramuscular injection, intraperitoneal injection, transdermally,orally, buccally, ocular routes, intravaginally, by inhalation, by depotinjections, or by implants.

In some embodiments, the compositions further comprise one or acombination of active agents chosen from: an anti-inflammatory compound,alpha-adrenergic agonist, antiarrhythmic compound, analgesic compound,and an anesthetic compound.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1 illustrates that PIF protects against lethal radiation. FIG. 1A:mice (C57BL/6, n=36) treated with PIF 2×/day for 14 days starting 2hours after 8Gy radiation exposure had 100% survival. Control mice(n=14) received radiation (PBS, vehicle) but no treatment, and developedARS and died by day 23. FIG. 1B: female mice (n=18, similar results inmales) were treated with PIF 2×/day (low, high dose: 0.75, 1.25 mg/kg)for 14 days starting 2 hours after 8Gy radiation exposure. Importantly,the PIF-treated group exhibited normal hematological indices, indicatingPIF's protective effect on hematopoiesis and immune function. Immuneprotection was evidenced by maintenance of lymphocyte and neutrophilnumbers. In addition, both hematocrit levels and platelet numbers werepreserved in the PIF-treated group. FIGS. 1C, 1D, and 1E: effects ofcontrol, low-dose, and high-dose PIF treatments, respectively, are shownon immune phenotype, RBCs, and platelets from day 0-19. Relevant figureabbreviations: WBC=white blood cells; NE=neutrophils; LY=lymphocytes;MO=monocytes; EO=eosinophils; BA=basophils; RBC=red blood cells;HB=hemoglobin; HCT=hematocrit; MCV=mean corpuscular volume; MCH=meancorpuscular hemoglobin; MCHC=mean corpuscular hemoglobin concentration;PLT=platelets; MPV=mean platelet volume.

FIG. 2A illustrates the survival curve after irradiation. Mice underwenta variety of doses of total body irradiation. The survival rate wasmonitored for 30 days post-irradiation. At 6-7Gy, all mice survived at30 days. FIG. 2B illustrates that PIF improves platelet count followingsub-lethal irradiation. Following exposure to 6Gy-8Gy irradiation doses,24-hour continuous treatment with PIF was initiated for 2 weeks,followed by 1 week post-therapy. The effect of the PIF treatment wascompared to a PBS control. Platelet count was determined, and is shownfor 6Gy, 7Gy, and 8Gy irradiation.

FIG. 3 illustrates that PIF enhances hematologic recovery aftersub-lethal irradiation. Mice were irradiated with a 6Gy dose. 1mg/kg/day of either PIF or PBS was administered continuously for twoweeks starting 24h after irradiation. The protocol of the experiment isdescribed in FIG. 3A. Follow-up of WBC reconstitution of the irradiatedmice is shown in FIG. 3B. WBC count 2 and 4 weeks after irradiation isshown in FIG. 3C. The percentages of lymphocytes and granulocytes 4weeks post-irradiation are shown in FIG. 3D. Results represent 2-3independent experiments. *P<0.05, **P<0.01.

FIG. 4 illustrates that PIF reduces inflammation and enhances B7H1expression after sub-lethal irradiation. Mice were irradiated witheither 6Gy or 7Gy doses. 0.75 mg/kg of either PIF or PBS wasadministered subcutaneously twice a day for three days, starting 24 hafter irradiation. The protocol of the experiment is described in FIG.4A. Levels of IL-1α and IL-2 in the serum of experimental mice weremeasured by FlowCytomix Multiplex kit for the 6Gy and 7Gy groups. Theresults are shown in FIGS. 4B and 4C, respectively. qPCR analyses ofiNOS and B7H1 mRNA expression in the colon were performed. The resultsare shown in FIGS. 4D and 4E, respectively. Results represent 2independent experiments. *P<0.05, **P<0.01.

FIG. 5A shows a sketch of the experiment. FIG. 5B illustrates that theeffect of sPIF was tested on colon crypt histology comparing the effectof sPIF initiated at 24 and 48 hours post-sub-lethal 6irradiation andgene expression (macro). FIG. 5C illustrates pictographs of sPIF'seffect as compared with PBS and normal mice. FIG. 5C illustrates that ascompared with PBS, the effect of sPIF at the two time point crypt depthwas significantly restored to that seen in the normal colon. FIGS. 5Dstatistical analysis demonstrates PIF reverses colon injury with nodifferences with normal mice. 5E illustrate qPCR B7H1 expressionincreased vs. PBS protection by increased B7H1 as compared with normaland PBS as well. Results represent two independent experiments. *P<0.05,**P<0.01.

FIG. 6 illustrates that PIF improves haematopoiesis after lethalirradiation and semi-allogeneic BMT. Mice were irradiated with a 10Gydose, which was followed by an semi-allogeneic BMT. 1 mg/kg/day ofeither PIF or PBS was administered continuously for two weeks starting24 h after irradiation. The protocol of the experiments is described inFIG. 6A. The WBC count of each group 3 weeks after irradiation andtransplantation is shown in FIG. 6B. The percentages of lymphocytes andgranulocytes 3 weeks post-irradiation are shown in FIG. 6C. Histologicalexamination of the femur bone for the cellularity level of the BM innormal, PBS-treated, and PIF-treated mice, as shown in FIGS. 6D, 6E, and6F, respectively. One representative picture out of 12 mice. The numberof fat cells in a 0.75 mm2 section of femur bone marrow is shown in FIG.6G. Results summarize 2 independent experiments (FIG. 6D). *P<0.05,****P<0.001.

FIG. 7 illustrates that BM pre-treated with PIF enhances hematologicrecovery after lethal irradiation and allogeneic BMT. Donor BM cellswere incubated with PIF for 2 h prior to transplantation. Mice wereirradiated with a 10Gy dose, followed by an allogeneic BMTtransplantation with the PIF-pre-treated BM graft. No additionaltreatments were given to the mice. The protocol of the experiments isdescribed in FIG. 7A. WBC and lymphocyte counts are shown 3 weeks (FIG.7B) and 4 weeks (FIG. 7C) after irradiation and transplantation. Resultsrepresent 3 independent experiments. *P<0.05. FIG. 3D illustrates thefemur bone. FIGS. 3E, 3F, and 3G illustrate that PIF enhancesmesenchymal stem cells' (MSCs') regulatory function. CFSE stained murinesplenocytes activated with anti-CD3 antibodies were cultured for fourdays (in a 50:1 ratio) with MSCs previously incubated (2 h) with PIF orcontrol. Cell proliferation was analyzed using flow cytometry. The graphin FIG. 3G shows % proliferating cells compared to control (activatedsplenocytes without MSCs), a summary of 3 experiments. H PIF promotesweight recovery after the transplant as compared to PBS, *P<0.05.

FIG. 8 illustrates that PIF shifts M1 macrophage differentiation to anM2-like phenotype. Peritoneal macrophages were cultured with GM-CSF (10ng/ml) and LPS (10 ng/ml) for M1 differentiation or with M-CSF (10ng/ml) and IL-4 (10 ng/ml) for M2 differentiation for 20 h in either thepresence or absence of PIF. qPCR analyses of iNOS (FIG. 8A), COX-2 (FIG.8B), and Arginase (FIG. 8C) mRNA expression of the differentiated cellswere performed. % of M1 macrophages gMFI of F480 (FIG. 8D) and CD11b(FIG. 8E) by FACS analysis are shown. Results represent 5-6 independentexperiments. *P<0.05, **P<0.01 ****P<0.001.

FIG. 9 illustrates FACS analyses of CD16/32 (FIG. 2A) and CD206 (FIG.2B). Peritoneal macrophages were cultured with GM-CSF (10 ng/ml) and LPS(10 ng/ml) for M1 differentiation or with M-CSF (10 ng/ml) and IL-4 (10ng/ml) for M2 differentiation for 20 h. Solid lines represent M1 anddashed lines represent M2 macrophages. One representative figure out of4 independent experiments.

FIG. 10 illustrates a heat map and cluster analysis of the global colongenome. Dark grey represents a relative decrease in expression ascompared to the pair of treatments or conditions. Lighter greyrepresents an increase in expression as compared to the treatment ofcondition. Column 1 illustrates a heat map of gene expression comparingexpression of genes in the presence of radiation+sPIF versus treatmentwith radiation alone. Column 2 depicts the comparative analysis ofrelative amount of gene expression from negative control (PBS) animalsversus those animals treated with radiation. Column 3 depicts therelative gene expression changes caused by radiation with sPIF ascompared to those mice untreated. Several pathways were affected. Thegenes most affected by sPIF and were genes associated with mitochondrialfunction, genes associated with response to stress and genes associatedwith protein-RNA interactions.

FIG. 11 illustrates data on the Differential Shift Assay analysis of PIFmutants for IDE, which is a binding partner for PIF. PIF mutants 1 and 3bind the Insulin-degrading enzyme (IDE).

FIG. 12 depicts data on the Differential Shift Assay analysis of PIFmutants for Kv1.3β, which is a binding partner for PIF. The bottom paneldepicts the change of Tm in 10 micromolar concentration of PIF mutants(left side versus 20 micromolar concentation of PIF (right side).

FIG. 13 illustrates the experimental protocol using PIF's injection fortesting the effect against lethal 50Gy radiation long term in rats. Itdepicts a flowchart of a protocol and dosing regimen describing when andhow organ transplant will be evaluated.

FIG. 14 illustrates the effect of PIF treatment following lethalradiation on body weight until 19 weeks of study in rats. PIF increasesrats weight at the end of the experiments vs PBS*<0.05. It depicts a oneway ANOVA plot of naive mice, mice irradiated with X-ray and thosetreated with PIF. n=9-12 mice. Holm Sidak post hoc test was performedwith a confidence interval of <0.05. PIF-treated animals that wereirradiated scored higher in body weight than those animals irradiatedalone.

FIG. 15 depicts a Kaplan Meier curve of PIF—treated animals as comparedto controls with a time period measured over 22 weeks of time. It showssurvival curve of PIF treated rats following lethal radiation. Nodifferences were found with controls.

-   Control, start: n=12, end: n=12-   X-Ray, start: n=12, end: n=10-   PIF, start: n=10, end: n=9

FIG. 16 depicts left ventricular wall thicknesses and diameters in mice.It depicts an image of cardiac function as examined by echography.

FIG. 17 depicts echocardiography results: left ventricular morphologynormalized to body weight. The results show that PIF ameloriates leftventricular hypertrophy at month 5. BW: body weight; AWTs: anterior wallthickness—systole; AWTd: anterior wall thickness—diastole, PWTs:posterior wall thickness—systole; PWTd: posterior wallthickness—diastole; SWTs: septal wall thickness—systole, SWTd: septalwall thickness—diastole. Two Way Repeated Measures ANOVA all pairwaisemultiple comparison. Holm Sidak post hoc test; vs control, p<0.05, # vsX-ray, p<0.05 significant interaction between group and time factors(n=9-12). It shows PIF effect of cardiac indices following lethalradiation. PIF improves both systolic and diastolic function as comparedto PBS wall thickness.

FIG. 18 depicts organ weights and lengths at month 5—body weight, heartweight, tibia length. One Way ANOVA on ranks, all pairwaise. Holm Sidakpost hoc test. * vs control, p<0.05# vs X-ray, p<0.05 n=9-12. It showsPIF effect on rat weight indices following lethal radiationdemonstrating increasing rats' weight vs PBS.

FIG. 19 depicts organ weights at month 5—pleural fluid, lung, kidney,liver and thymus weight One Way ANOVA on ranks, all pairwaise. HolmSidak post hoc test, * vs control, p<0.05# vs X-ray, p<0.05, n=9-12. Itshows PIF effect on rat organ weight following lethal radiation. PIFpromotes kidney growth.

FIG. 20 depicts the effects of PIF on adrenal cell cultures as comparedto untreated cells. BAC cells treated with PIF have an increasedviability in culture when pre-exposed to PIF shows PIF effect on bovineadrenal cells (primary) viability, apoptosis and proliferation.

FIG. 21 shows the PIF effect on cortisol secretion by bovine adrenalcells (primary); the effect of PIF on BAC adrenal cells as it relates tobasal cortisol secretion. Cortisol levels increase when PIF is exposedto cells in culture.

FIG. 22 shows effect of PIF on INS-1 (rat islet insulin producing cells)in culture. Cell viability is increased in culture as compared to INS-1cells left untreated. Doses of PIF at 0.01, 0.1, and 1.0 micrograms/mLof culture medium were used in the treatment. It shows that PIF promotesinsulinoma cells viability, as well as viability.

DETAILED DESCRIPTION

Acute radiation syndrome (ARS), also known as radiation sickness,develops after whole-body or partial-body high-dose irradiation.Radiation may cause complete destruction of the bone marrow, damage tothe mucosal barrier and crypts of the gastrointestinal (GI) tract, skinburns, and central nervous system injury leading to irreversibleneurologic and cardiovascular damage, and ultimately to death. Radiationis particular harmful to rapid turnover cells—such as hematopoieticcells—with lymphocytes being the first sub-lineage to be depleted.Although individual organ damage can be monitored, a more suitable viewfor ARS is the concept of multiple organ dysfunction syndrome caused bya systemic inflammatory response. A number of reports show thatradiation-induced production of proinflammatory cytokines contributes toradiotherapy-associated disorders in the blood and peripheral lymphoidtissues.

Given the complexity of ionizing radiation-induced injury, effective ARStherapies are lacking. The current clinical approach for treatment is toinhibit the production of inflammatory mediators and suppress theinitiation of the inflammatory response. Current management includesblood transfusion, fluid and electrolytes administration, antibiotics,and antiviral therapy. These treatments cause generalizedimmunosuppression and place patients in danger of opportunisticinfections. Patients with cytopenia receive granulocytecolony-stimulating factor or granulocyte macrophage colony-stimulatingfactor to re-populate the immune system from residual hematopoieticprogenitor cells effective following low grade radiation. Non-respondersand following lethal radiation require hematopoietic stem-cellstransplantation (HSCT). However, such transplantation frequently leadsto deleterious graft vs host disease (GVHD) coupled with impaired graftvs. leukemia (GVL) effect due to under or over use of immune suppressivedrugs . ARS currently has only limited countermeasures.

Bone marrow transplantation (BMT) may be used as a treatment forhematological malignancies and inherited blood cell disorders, such as,but not limited to, lymphomas, lymphocytic leukemias, myeloma, leukemia,anemia, hemophilia, thalassemias, sickle cell disease, multiplesclerosis, scleroderma, myelodysplastic syndromes and myeloproliferativediseases. There are two types of bone marrow transplantation: autologous(self) and allogeneic (donor).

The transplantation of organs, tissue, or cells other than bone marrowmay also be used to treat various physiological deficiencies of thetransplant recipient, including the dysfunction of multiple organs thatmay occur with ARS. Such organs, tissue, or cells may include, but arenot limited to, the skin, brain, heart, lungs, kidneys, gastrointestinaltract, spleen, liver, pancreas, pancreatic islet cells, adrenal glands,and combinations thereof.

Autologous transplantation, allogeneic transplantation, semi-allogeneictransplantation, or xenotransplantation may be used if the subjectrequires transplantation. In the case of xenotransplantation, forexample, islet cells or other cells, organs, or tissues from a porcinedonor may be transplanted due to the current major shortage of humanorgans available for transplantation. The perfect matching of organs,tissue, or cells for transplantation is a major quest to preventtransplant rejection or, in the case of bone marrow, failure to engraftor, conversely, the development of graft-versus-host disease (GVHD).However, current therapies are limited, and the failure of the graft andother major complications still occur. The introduction ofimmunosuppressive agents has significantly advanced the field oftransplantation, allowing patients to recover long-term; the patients,however, require life-long immunosuppressive therapy, which isassociated with toxic side effects.

An autologous transplant may be possible if the disease afflicting thetarget organ, tissue, or cell is in remission, or if the condition beingtreated does not involve the target organ, tissue, or cell. Inautologous transplantation, including BMT, the tissue is extracted fromthe patient prior to transplantation and may be “purged” to removelingering malignant cells (if the disease has afflicted the targetorgan, tissue, or cell). After the patient undergoes chemotherapy orradiation, the stem cells are transplanted back into the patient.Autologous transplant allows the patient to receive high doses ofchemotherapy and radiation. High doses of chemotherapy or radiationtherapy with bone marrow or peripheral blood transplants, for example,have improved cure rates for leukemia and lymphoma. Once the patient hasundergone chemotherapy or radiation, the patient may have a limitedability to produce blood cells. An autologous transplant would allow thepatient to “jump start” the production of blood cells and platelets.

Mammalian pregnancy is a unique physiological event in which thematernal immune system interacts with the fetus in a very efficientmanner that is beneficial for both parties. Pregnancy is an immuneparadox, displaying no graft vs. host or host vs. graft effect. Thefactors involved in this phenomenon are not yet fully elucidated,although they have been extensively studied. The novel embryo-derivedfactor, PreImplantation Factor (PIF-1), may cause immune tolerance ofpregnancy by creating maternal recognition of pregnancy shortly afterfertilization.

To transpose PIF therapeutic utility outside pregnancy, a synthetic PIFanalog (sPIF) of 15 amino acids (MVRIKPGSANKPSDD) mimicking the nativepeptide activity was generated (and upgraded to cGMP grade). Thisenabled detailed examination of sPIF effect in preclinical models ofautoimmunity, transplant and acute radiation syndrome showing anintegrated local and systemic efficacy. Studies examining sPIF's effecton human immune cells and determining crucial elements of its local andsystemic mechanism of action were successful. sPIF efficacy to preventand reverse in semi-allogenic graft vs. host model while preservingbeneficial graft vs. leukemia was documented. FDA-mandated comprehensivetoxicology studies demonstrated a high safety profile (mice and dogs) at(supra-physiologic (400-4000× human) doses coupled with shortcirculating half-life (Covance). The FDA has granted FAST TRACKdesignation to sPIF to conduct University-sponsored clinical trials foran immune orphan disorder. Synthetic PIF-1 (sPIF) replicated the nativepeptide's effect and exerted potent immune modulatory effects onactivated peripheral blood mononuclear cell (PBMC) proliferation andcytokine secretion, acting through novel sites on PBMC and having aneffect which is distinct from known immunosuppressive drugs. Therefore,PIF may prevent the development of GVHD while allowing graft vs.leukemia to be effective. Without wishing to be bound by theory, PIF maybind to transplanted bone marrow stem cells and help them becomeintegrated quickly and start producing the normal blood cells andplatelets.

A transplant is ideally conducted with a donor organ, tissue, or cellthat perfectly matches the recipient; still, about 70% of patientsdevelop GVHD of various intensities. Therefore, transplant patientsreceive immunosuppressive therapy post-transplant, and possibly for theremainder of their lives. Beyond acute GVHD, chronic GVHD and rejectionare additional serious complications. Those conditions, however, aremore difficult to treat and have high morbidity and mortality whencompared to acute GVHD. Therefore, the use of PIF may enable autologoustransplantation, allogeneic transplantation, semi-allogeneictransplantation, or xenotransplantation, thereby improving engraftment,preventing GVHD or organ rejection long-term.

Clinically, the major problem in transplantation occurs during the longtime until the transplanted cells become functional. During that timethe patient often gets serious infections and may die. PIF addresses thefundamentals of inflammation regardless of the origin of injury or whichorgan is targeted. PIF administration may reduce the engraftment timeperiod and engraftment-associated inflammation following transplant. PIFmay lead to successful transplant engraftment. Without wishing to bebound by theory, it is believed that similar to PIF's activity inpregnancy, PIF outside pregnancy facilitates engraftment and allows thenewly incorporated organ, tissue, or cells to become functional.Furthermore, administration of PIF may allow for full weight recovery inmice receiving an autologous transplant, which is comparable to that ofnaïve mice.

sPIF may prevent deleterious GVHD development while preserving thebeneficial GVL effect, as shown in a murine allogeneic bone marrowtransplantation (BMT) model. In that model, short-term sPIF therapy ledto four-month efficacy protection against dermatitis, hepatitis, andcolon ulceration coupled with decreased pro-inflammatory hepaticcytokines and chemokines genes and circulating pro-inflammatory IL17alevels. sPIF has also been shown to promote syngeneic BMT.

sPIF regulates global immunity, and targets naive monocytes/macrophagesacting on activated immunity to block mixed lymphocyte reaction (MLR),proliferation leading to a preferential TH2 cytokine bias whilepreserving the necessary TH1 response. In murine macrophages, increasedB7H1 (ligated to PD-1 on T-cells) and decreased iNOS gene (NOS2, nitricoxide synthase) expression have been noted (Azar et al., 2013). sPIFtargets protein-di-isomerase/Thioredoxin (PDI/T) and heat shock proteins(HSPs) to reduce oxidative stress and prevent protein misfolding,thereby providing important insight into the observed PIF protection inpreclinical models. Observations both in vitro and in preclinical modelssupport the view that sPIF may be effective in protecting against ARS.

Before the present compositions and methods are described, it is to beunderstood that this invention is not limited to the particularprocesses, compositions, or methodologies described, as these may vary.It is also to be understood that the terminology used in the descriptionis for the purpose of describing the particular versions or embodimentsonly, and is not intended to limit the scope of the present inventionwhich will be limited only by the appended claims. Unless definedotherwise, all technical and scientific terms used herein have the samemeanings as commonly understood by one of ordinary skill in the art.Although any methods and materials similar or equivalent to thosedescribed herein can be used in the practice or testing of embodimentsof the present invention, the preferred methods, devices, and materialsare now described. All publications mentioned herein are incorporated byreference in their entirety. Nothing herein is to be construed as anadmission that the invention is not entitled to antedate such disclosureby virtue of prior invention.

It must also be noted that as used herein and in the appended claims,the singular forms “a”, “an”, and “the” include plural reference unlessthe context clearly dictates otherwise. Thus, for example, reference toa “peptide” is a reference to one or more peptides and equivalentsthereof known to those skilled in the art, and so forth.

As used herein, the term “about” means plus or minus 10% of thenumerical value of the number with which it is being used. Therefore,about 50% means in the range of 40%-60%.

“Administering” when used in conjunction with a therapeutic means toadminister a therapeutic directly into or onto a target organ, tissue orcell or to administer a therapeutic to a patient, whereby thetherapeutic positively impacts the organ, tissue or cell to which it istargeted. Thus, as used herein, the term “administering”, when used inconjunction with pre-implantation factor (PIF), can include, but is notlimited to, providing PIF into or onto the target organ, tissue or cell;providing PIF systemically to a patient by, e.g., intravenous injectionwhereby the therapeutic reaches the target organ, tissue or cell;providing PIF in the form of the encoding sequence thereof to the targettissue (e.g., by so-called gene-therapy techniques). “Administering” maybe accomplished by parenteral, oral or topical administration, or bysuch methods in combination with other known techniques.

The terms “animal,” “patient,” and “subject” as used herein include, butare not limited to, humans and non-human vertebrates such as wild,domestic and farm animals. In some embodiments, the terms “animal,”“patient,” and “subject” may refer to humans. In some embodiments, theterms “animal,” “patient,” and “subject” may refer to non-human mammals.In some embodiments, the terms “animal,” “patient,” and “subject” mayrefer to any or combination of: dogs, cats, pigs, cows, horses, goats,sheep or other domesticated non-human mammals. In some embodiments, thesubject is a human patient that has been diagnosed or is suspected ofhaving a malignant form of cancer. In some embodiments, the subject is ahuman patient that has been diagnosed or is suspected of having organfailure. In some embodiments, the subject is a human patient that hasbeen diagnosed or is suspected of having liver failure, kidney failure,any disease associated with an imbalance of cortisol levels, juvenile oradult diabetes (type I or II), or heart failure. In some embodiments,the subject is a human patient that has been identified as requiring orsuspected of requiring an Islet cell transplant, a kidney transplant, oradrenal cell transplant, a blood cell transplant, a bone marrowtransplant, or a heart transplant.

“Immune-modulating” refers to the ability of a compound of the presentinvention to alter (modulate) one or more aspects of the immune system.The immune system functions to protect the organism from infection andfrom foreign antigens by cellular and humoral mechanisms involvinglymphocytes, macrophages, and other antigen-presenting cells thatregulate each other by means of multiple cell-cell interactions and byelaborating soluble factors, including lymphokines and antibodies, thathave autocrine, paracrine, and endocrine effects on immune cells.

The term “improves” is used to convey that the present invention changeseither the appearance, form, characteristics and/or the physicalattributes of the subject, organ, tissue or cell to which it is beingprovided, applied or administered. For example, the change in form maybe demonstrated by any of the following alone or in combination: adecrease in one or more symptoms of ARS; increased engraftment oftransplanted organs, tissues, or cells; increased acceptance oftransplanted organs, tissues, or cells; reduction of host immuneresponse to graft associated with autologous transplant, allogeneictransplant, semi-allogeneic transplant, or xenotransplant; increasedgraft v. leukemia; increase in graft v. leukemia with no or with minimalgraft v. host disease; reduction or elimination of the need for immunesuppressive agents; and faster recovery from chemotherapy and radiationtherapy.

The term “inhibiting” includes the administration of a compound of thepresent invention to prevent the onset of the symptoms, alleviating thesymptoms, or eliminating the disease, condition or disorder.

As used herein, the terms “peptide,” “polypeptide” and “protein” areused interchangeably and refer to two or more amino acids covalentlylinked by an amide bond or non-amide equivalent. The peptides of theinvention can be of any length. For example, the peptides can have fromabout two to about 100 or more residues, such as, 5 to 12, 12 to 15, 15to 18, 18 to 25, 25 to 50, 50 to 75, 75 to 100, or more in length.Preferably, peptides are from about 2 to about 18 residues. The peptidesof the invention include l- and d-isomers, and combinations of l- andd-isomers. The peptides can include modifications typically associatedwith post-translational processing of proteins, for example, cyclization(e.g., disulfide or amide bond), phosphorylation, glycosylation,carboxylation, ubiquitination, myristylation, or lipidation.

By “pharmaceutically acceptable,” it is meant the carrier, diluent orexcipient must be compatible with the other ingredients of theformulation or composition and not deleterious to the recipient thereof.

Unless otherwise indicated, the term “bone marrow” means that flexibletissue found in the hollow interior of bones, consisting of the redmarrow and yellow marrow, and containing stem cells.

As used herein, the term “therapeutic” means an agent utilized to treat,combat, ameliorate, prevent or improve an unwanted condition or diseaseof a patient. In part, embodiments of the present invention are directedto decreasing one or more symptoms of ARS, an increase in acceptance oftransplanted organs, tissues, or cells in autologous transplants,allogeneic transplants, semi-allogeneic transplants, or xenotransplants,and/or a decrease in the rejection of organs, tissues, or cells inautologous transplants, allogeneic transplants, semi-allogeneictransplants, or xenotransplants.

A “therapeutically effective amount” or “effective amount” of acomposition (e.g, a PIF peptide) is a predetermined amount calculated toachieve the desired effect, i.e., to improve, increase, or allow theacceptance of organs, tissues, or cells in autologous transplantation,allogeneic transplantation, semi-allogeneic transplantation, orxenotransplantation, and/or to decrease one or more symptoms of ARS orincrease the viability of donor organs, tissues, or cell before they aretransplanted. The activity contemplated by the present methods includesboth medical therapeutic and/or prophylactic treatment, as appropriate.The specific dose of a compound administered according to this inventionto obtain therapeutic and/or prophylactic effects will, of course, bedetermined by the particular circumstances surrounding the case,including, for example, the compound administered, the route ofadministration, and the condition being treated. The compounds areeffective over a wide dosage range and, for example, dosages per daywill normally fall within the range of from 0.001 to 10 mg/kg, moreusually in the range of from 0.01 to 1 mg/kg. In some embodiments, thetherapeutically effective dose of PIF or PIF analog or peptide is about0.1 mg/kg, 0.2 mg/kg, 0.3 mg/kg, 0.4 mg/kg, 0.5 mg/kg, 0.6 mg/kg, 0.7mg/kg, 0.8 mg/kg, 0.9 mg/kg, and 1 mg/kg. However, it will be understoodthat the effective amount administered will be determined by thephysician in the light of the relevant circumstances including thecondition to be treated, the choice of compound to be administered, andthe chosen route of administration, and therefore the above dosageranges are not intended to limit the scope of the invention in any way.A therapeutically effective amount of compound of embodiments of thisinvention is typically an amount such that when it is administered in aphysiologically tolerable excipient composition, it is sufficient toachieve an effective systemic concentration or local concentration inthe tissue.

The terms “treat,” “treated,” or “treating” as used herein refers toboth therapeutic treatment and prophylactic or preventative measures,wherein the object is to prevent or slow down (lessen) an undesiredphysiological condition, disorder or disease, or to obtain beneficial ordesired clinical results. For the purposes of this invention, beneficialor desired clinical results include, but are not limited to, alleviationof symptoms; diminishment of the extent of the condition, disorder ordisease; stabilization (i.e., not worsening) of the state of thecondition, disorder or disease; delay in onset or slowing of theprogression of the condition, disorder or disease; amelioration of thecondition, disorder or disease state; and remission (whether partial ortotal), whether detectable or undetectable, or enhancement orimprovement of the condition, disorder or disease. Treatment includeseliciting a clinically significant response without excessive levels ofside effects. Treatment also includes prolonging survival as compared toexpected survival if not receiving treatment.

Generally speaking, the term “tissue” refers to any aggregation ofsimilarly specialized cells which are united in the performance of aparticular function.

This application describes compounds. Without being bound by anyparticular theory, the compounds described herein act as agonists ofPIF-mediated signal transduction via the receptor or receptors of PIF.Thus, these compounds modulate signaling pathways that providesignificant therapeutic benefit in the treatment of, but not limited to,acute radiation syndrome and delayed effects of acute radiationexposure. The compounds of the present disclosure may exist inunsolvated forms as well as solvated forms, including hydrated forms.The compounds of the present disclosure also are capable of forming bothpharmaceutically acceptable salts, including but not limited to acidaddition and/or base addition salts. Furthermore, compounds of thepresent disclosure may exist in various solid states including anamorphous form (non-crystalline form), and in the form of clathrates,prodrugs, polymorphs, bio-hydrolyzable esters, racemic mixtures,non-racemic mixtures, or as purified stereoisomers including, but notlimited to, optically pure enantiomers and diastereomers. In general,all of these forms can be used as an alternative form to the free baseor free acid forms of the compounds, as described above and are intendedto be encompassed within the scope of the present disclosure.

A “polymorph” refers to solid crystalline forms of a compound. Differentpolymorphs of the same compound can exhibit different physical, chemicaland/or spectroscopic properties. Different physical properties include,but are not limited to stability (e.g., to heat or light),compressibility and density (important in formulation and productmanufacturing), and dissolution rates (which can affectbioavailability). Different physical properties of polymorphs can affecttheir processing. In some embodiments, the pharmaceutical compositioncomprises at least one polymorph of any fo the compositions disclosedherein.

As noted above, the compounds of the present disclosure can beadministered, inter alia, as pharmaceutically acceptable salts, esters,amides or prodrugs. The term “salts” refers to inorganic and organicsalts of compounds of the present disclosure. The salts can be preparedin situ during the final isolation and purification of a compound, or byseparately reacting a purified compound in its free base or acid formwith a suitable organic or inorganic base or acid and isolating the saltthus formed. Representative salts include the hydrobromide,hydrochloride, sulfate, bisulfate, nitrate, acetate, oxalate,palmitiate, stearate, laurate, borate, benzoate, lactate, phosphate,tosylate, citrate, maleate, fumarate, succinate, tartrate, naphthylate,mesylate, glucoheptonate, lactobionate, and laurylsulphonate salts, andthe like. The salts may include cations based on the alkali and alkalineearth metals, such as sodium, lithium, potassium, calcium, magnesium,and the like, as well as non-toxic ammonium, quaternary ammonium, andamine cations including, but not limited to, ammonium,tetramethylammonium, tetraethylammonium, methylamine, dimethylamine,trimethylamine, triethylamine, ethylamine, and the like. See, forexample, S. M. Berge, et al., “Pharmaceutical Salts,” J Pharm Sci, 66:1-19 (1977). The term “salt” refers to acidic salts formed withinorganic and/or organic acids, as well as basic salts formed withinorganic and/or organic bases. Examples of these acids and bases arewell known to those of ordinary skill in the art. Such acid additionsalts will normally be pharmaceutically acceptable although salts ofnon-pharmaceutically acceptable acids may be of utility in thepreparation and purification of the compound in question. Salts includethose formed from hydrochloric, hydrobromic, sulphuric, phosphoric,citric, tartaric, lactic, pyruvic, acetic, succinic, fumaric, maleic,methanesulphonic and benzenesulphonic acids.

In some embodiments, salts of the compositions comprising either a PIFor PIF analog or PIF mutant may be formed by reacting the free base, ora salt, enantiomer or racemate thereof, with one or more equivalents ofthe appropriate acid. In some embodiments, pharmaceutical acceptablesalts of the present disclosure refer to analogs having at least onebasic group or at least one basic radical. In some embodiments,pharmaceutical acceptable salts of the present disclosure comprise afree amino group, a free guanidino group, a pyrazinyl radical, or apyridyl radical that forms acid addition salts. In some embodiments, thepharmaceutical acceptable salts of the present disclosure refer toanalogs that are acid addition salts of the subject compounds with (forexample) inorganic acids, such as hydrochloric acid, sulfuric acid or aphosphoric acid, or with suitable organic carboxylic or sulfonic acids,for example aliphatic mono- or di-carboxylic acids, such astrifluoroacetic acid, acetic acid, propionic acid, glycolic acid,succinic acid, maleic acid, fumaric acid, hydroxymaleic acid, malicacid, tartaric acid, citric acid or oxalic acid, or amino acids such asarginine or lysine, aromatic carboxylic acids, such as benzoic acid,2-phenoxy-benzoic acid, 2-acetoxybenzoic acid, salicylic acid,4-aminosalicylic acid, aromatic-aliphatic carboxylic acids, such asmandelic acid or cinnamic acid, heteroaromatic carboxylic acids, such asnicotinic acid or isonicotinic acid, aliphatic sulfonic acids, such asmethane-, ethane- or 2-hydroxyethane-sulfonic acid, or aromatic sulfonicacids, for example benzene-, p-toluene- or naphthalene-2-sulfonic acid.When several basic groups are present mono- or poly-acid addition saltsmay be formed. The reaction may be carried out in a solvent or medium inwhich the salt is insoluble or in a solvent in which the salt issoluble, for example, water, dioxane, ethanol, tetrahydrofuran ordiethyl ether, or a mixture of solvents, which may be removed in vacuoor by freeze drying. The reaction may also be a metathetical process orit may be carried out on an ion exchange resin. In some embodiments, thesalts may be those that are physiologically tolerated by a patient.Salts according to the present disclosure may be found in theiranhydrous form or as in hydrated crystalline form (i.e., complexed orcrystallized with one or more molecules of water).

Examples of pharmaceutically acceptable esters of the compounds of thepresent disclosure include C₁-C₈ alkyl esters. Acceptable esters alsoinclude C₅-C₇ cycloalkyl esters, as well as arylalkyl esters such asbenzyl. C₁-C₄ alkyl esters are commonly used. Esters of compounds of thepresent disclosure may be prepared according to methods that are wellknown in the art. Examples of pharmaceutically acceptable amides of thecompounds of the present disclosure include amides derived from ammonia,primary C₁-C₈ alkyl amines, and secondary C₁-C₈ dialkyl amines. In thecase of secondary amines, the amine may also be in the form of a 5 or 6membered heterocycloalkyl group containing at least one nitrogen atom.Amides derived from ammonia, C₁-C₃ primary alkyl amines and C₁-C₂dialkyl secondary amines are commonly used. Amides of the compounds ofthe present disclosure may be prepared according to methods well knownto those skilled in the art.

As used herein, “conservative” amino acid substitutions may be definedas set out in Tables A, B, or C below. The PIF compounds of thedisclosure include those wherein conservative substitutions (from eithernucleic acid or amino acid sequences) have been introduced bymodification of polynucleotides encoding polypeptides of the disclosure.Amino acids can be classified according to physical properties andcontribution to secondary and tertiary protein structure. A conservativesubstitution is recognized in the art as a substitution of one aminoacid for another amino acid that has similar properties. In someembodiments, the conservative substitution is recognized in the art as asubstitution of one nucleic acid for another nucleic acid that hassimilar properties, or, when encoded, has similar binding affinities.Exemplary conservative substitutions are set out in Table 1.

TABLE 1 Conservative Substitutions I Side Chain Characteristics AminoAcid Aliphatic Non-polar G A P I L V F Polar - uncharged C S T M N QPolar - charged D E K R Aromatic H F W Y Other N Q D E

Alternately, conservative amino acids can be grouped as described inLehninger, (Biochemistry, Second Edition; Worth Publishers, Inc. NY,N.Y. (1975), pp. 71-77) as set forth in Table 2.

TABLE 2 Conservative Substitutions II Side Chain Characteristic AminoAcid Non-polar (hydrophobic) Aliphatic: A L I V P Aromatic: F W YSulfur-containing: M Borderline: G Y Uncharged-polar Hydroxyl: S T YAmides: N Q Sulfhydryl: C Borderline: G Y Positively Charged (Basic): KR H Negatively Charged (Acidic): D E

Alternately, exemplary conservative substitutions are set out in Table3.

TABLE 3 Conservative Substitutions III Original Residue ExemplarySubstitution Ala (A) Val Leu Ile Met Arg (R) Lys His Asn (N) Gln Asp (D)Glu Cys (C) Ser Thr Gln (Q) Asn Glu (E) Asp Gly (G) Ala Val Leu Pro His(H) Lys Arg Ile (I) Leu Val Met Ala Phe Leu (L) Ile Val Met Ala Phe Lys(K) Arg His Met (M) Leu Ile Val Ala Phe (F) Trp Tyr Ile Pro (P) Gly AlaVal Leu Ile Ser (S) Thr Thr (T) Ser Trp (W) Tyr Phe Ile Tyr (Y) Trp PheThr Ser Val (V) Ile Leu Met Ala

As used herein, the terms “peptide,” “polypeptide” and “protein” areused interchangeably and refer to two or more amino acids covalentlylinked by an amide bond or non-amide equivalent. The peptides of thedisclosure can be of any length. For example, the peptides can have fromabout two to about 100 or more residues, such as, 5 to 12, 12 to 15, 15to 18, 18 to 25,25 to 50,50 to 75,75 to 100, or more in length.Preferably, peptides are from about 2 to about 18 residues in length.The peptides of the disclosure also include l- and d-isomers, andcombinations of l- and d-isomers. The peptides can include modificationstypically associated with posttranslational processing of proteins, forexample, cyclization (e.g., disulfide or amide bond), phosphorylation,glycosylation, carboxylation, ubiquitination, myristylation, orlipidation. In some embodiments, the compositions or pharmaceuticalcompositions of the disclosure relate to analogs of any PIF sequence setforth in Table 1 that share no less than about 70%, about 75%, about79%, about 80%, about 85%, about 86%, about 87%, about 90%, about 93%,about 94% about 95%, about 96%, about 97%, about 98%, about 99% homologywith any one or combination of PIF sequences set forth in Table 1. Insome embodiments, PIF or PIF peptide may refer to an amino acid sequenceselected from SEQ ID NOs: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13 ,14,15, 16 ,17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29 or afunctional fragment thereof that is about 70%, 75%, 80%, 85%, 90%, 91%,92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% homologous to any such aminoacid sequence. In some embodiments, PIF may refer to an amino acidsequence comprising, consisting essentially of, or consisting of asequence that is at least 70%, 75%, 80%, 85%, 86%, 87%, 90%, 91%, 92%,93%, 94%, 95%, 96%, 97%, 98%, or 99% homologous to SEQ ID. NO: 20. Insome embodiments, PIF may refer to an amino acid sequence comprising,consisting essentially of, or consisting of a sequence that is at least70%, 75%, 80%, 85%, 86%, 87%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%,98%, or 99% homologous to SEQ ID. NO: 21. In some embodiments, PIF mayrefer to an amino acid sequence comprising, consisting essentially of,or consisting of a sequence that is at least 70%, 75%, 80%, 85%, 86%,87%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% homologous toSEQ ID. NO: 22. In some embodiments, PIF may refer to an amino acidsequence comprising, consisting essentially of, or consisting of asequence that is at least 70%, 75%, 80%, 85%, 86%, 87%, 90%, 91%, 92%,93%, 94%, 95%, 96%, 97%, 98%, or 99% homologous to SEQ ID. NO: 23. Insome embodiments, PIF may refer to an amino acid sequence comprising,consisting essentially of, or consisting of a sequence that is at least70%, 75%, 80%, 85%, 86%, 87%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%,98%, or 99% homologous to SEQ ID. NO: 24. In some embodiments, PIF mayrefer to an amino acid sequence comprising, consisting essentially of,or consisting of a sequence that is at least 70%, 75%, 80%, 85%, 86%,87%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% homologous toSEQ ID. NO: 25. In some embodiments, PIF may refer to an amino acidsequence comprising, consisting essentially of, or consisting of asequence that is at least 70%, 75%, 80%, 85%, 86%, 87%, 90%, 91%, 92%,93%, 94%, 95%, 96%, 97%, 98%, or 99% homologous to SEQ ID. NO: 26. Insome embodiments, PIF may refer to an amino acid sequence comprising,consisting essentially of, or consisting of a sequence that is at least70%, 75%, 80%, 85%, 86%, 87%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%,98%, or 99% homologous to SEQ ID. NO: 27. In some embodiments, PIF mayrefer to an amino acid sequence comprising, consisting essentially of,or consisting of a sequence that is at least 70%, 75%, 80%, 85%, 86%,87%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% homologous toSEQ ID. NO: 28. In some embodiments, PIF may refer to an amino acidsequence comprising, consisting essentially of, or consisting of asequence that is at least 70%, 75%, 80%, 85%, 86%, 87%, 90%, 91%, 92%,93%, 94%, 95%, 96%, 97%, 98%, or 99% homologous to SEQ ID. NO: 29. Insome embodiments, the PIF mutant comprises a sequence selected from:XVZIKPGSANKPSD, XVZIKPGSANKPS XVZIKPGSANKP XVZIKPGSANK XVZIKPGSAN,XVZIKPGSA, XVZIKPGS, XVZIKPG, XVZIKP, XVZIK, XVZI, XVZ wherein X is anon-natural amino acid or a naturally occurring amino acid. In someembodiments, the PIF mutant comprises a sequence selected from:XVZIKPGSANKPSD, XVZIKPGSANKPS XVZIKPGSANKP XVZIKPGSANK XVZIKPGSAN,XVZIKPGSA, XVZIKPGS, XVZIKPG, XVZIKP, XVZIK, XVZI, XVZ wherein X is anon-natural amino acid or a naturally occurring amino acid except that Xis not methionine if Z is arginine, and Z is not arginine if X ismethionine. In some embodiments, the PIF analog or mutant is syntheticor synthetically made.

Peptides disclosed herein further include compounds having amino acidstructural and functional analogs, for example, peptidomimetics havingsynthetic or non-natural amino acids (such as a norleucine) or aminoacid analogues or non-natural side chains, so long as the mimetic sharesone or more functions or activities of compounds of the disclosure. Thecompounds of the disclosure therefore include “mimetic” and“peptidomimetic” forms. As used herein, a “non-natural side chain” is amodified or synthetic chain of atoms joined by covalent bond to theα-carbon atom, β-carbon atom, or γ-carbon atom which does not make upthe backbone of the polypeptide chain of amino acids. The peptideanalogs may comprise one or a combination of non-natural amino-acidschosen from: norvaline, tert-butylglycine, phenylglycine, He,7-azatryptophan, 4-fluorophenylalanine, N-methyl-methionine,N-methyl-valine, N-methyl-alanine, sarcosine,N-methyl-tert-butylglycine, N-methyl-leucine, N-methyl-phenylglycine,N-methyl-isoleucine, N-methyl-tryptophan, N-methyl-7-azatryptophan,N-methyl-phenylalanine, N-methyl-4-fluorophenyl alanine,N-methyl-threonine, N-methyl-tyrosine, N-methyl-valine, N-methyl-lysine,homocysteine, and Tyr; Xaa2 is absent, or an amino acid selected fromthe group consisting of Ala, D-Ala, N-methyl-alanine, Glu,N-methyl-glutamate, D-Glu, Gly, sarcosine, norleucine, Lys, D-Lys, Asn,D-Asn, D-Glu, Arg, D-Arg, Phe, D-Phe, N-methyl-phenylalanine, Gin,D-Gln, Asp, D-Asp, Ser, D-Ser, N-methyl-serine, Thr, D-Thr,N-methyl-threonine, D-Pro D-Leu, N-methyl-leucine, D-Ile,N-methyl-isoleucine, D-Val, N-methyl-valine, tert-butylglycine,D-tert-butylglycine, N-methyl-tert-butylglycine, Trp, D-Trp,N-methyl-tryptophan, D-Tyr, N-methyl-tyrosine,1-aminocyclopropanecarboxylic acid, 1-aminocyclobutanecarboxylic acid,1-aminocyclopentanecarboxylic acid, 1-aminocyclohexanecarboxylic acid,4-aminotetrahydro-2H-pyran-4-carboxylic acid, aminoisobutyric acid,(5)-2-amino-3-(1H-tetrazol-5-yl)propanoic acid, Glu, Gly,N-methyl-glutamate, 2-amino pentanoic acid, 2-amino hexanoic acid,2-amino heptanoic acid, 2-amino octanoic acid, 2-amino nonanoic acid,2-amino decanoic acid, 2-amino undecanoic acid, 2-amino dodecanoic acid,octylglycine, tranexamic acid, aminovaleric acid, and2-(2-aminoethoxy)acetic acid. The natural side chain, or R group, of analanine is a methyl group. In some embodiments, the non-natural sidechain of the composition is a methyl group in which one or more of thehydrogen atoms is replaced by a deuterium atom. Non-natural side chainsare disclosed in the art in the following publications: WO/2013/172954,WO2013123267, WO/2014/071241, WO/2014/138429, WO/2013/050615,WO/2013/050616, WO/2012/166559, US Application No. 20150094457, Ma, Z.,and Hartman, M. C. (2012). In Vitro Selection of Unnatural CyclicPeptide Libraries via mRNA Display. In J. A. Douthwaite & R. H. Jackson(Eds.), Ribosome Display and Related Technologies: Methods and Protocols(pp. 367-390). Springer New York., all of which are incorporated byreference in their entireties.

The terms “mimetic,” “peptide mimetic” and “peptidomimetic” are usedinterchangeably herein, and generally refer to a peptide, partialpeptide or non-peptide molecule that mimics the tertiary bindingstructure or activity of a selected native peptide or protein functionaldomain (e.g., binding motif or active site). These peptide mimeticsinclude recombinantly or chemically modified peptides, as well asnon-peptide agents such as small molecule drug mimetics, as furtherdescribed below. The term “analog” refers to any polypeptide comprisingat least one a-amino acid and at least one non-native amino acidresidue, wherein the polypeptide is structurally similar to a naturallyoccurring full-length PIF protein and shares the biochemical orbiological activity of the naturally occurring full-length protein uponwhich the analog is based. In some embodiments, the compositions,pharmaceutical compositions and kits comprise a peptide or peptidomimeicsharing share no less than about 70%, about 75%, about 79%, about 80%,about 85%, about 86%, about 87%, about 90%, about 93%, about 94% about95%, about 96%, about 97%, about 98%, about 99% homology with any one orcombination of PIF sequences set forth in Table 4; and wherein one or aplurality of amino acid residues is a non-natural amino acid residue oran amino acid residue with a non-natural sidechain. In some embodiments,peptide or peptide mimetics are provided, wherein a loop is formedbetween two cysteine residues. In some embodiments, the peptidomimeticmay have many similarities to natural peptides, such as: amino acid sidechains that are not found among the known 20 proteinogenic amino acids,non-peptide-based linkers used to effect cyclization between the ends orinternal portions of the molecule, substitutions of the amide bondhydrogen moiety by methyl groups (N-methylation) or other alkyl groups,replacement of a peptide bond with a chemical group or bond that isresistant to chemical or enzymatic treatments, N- and C-terminalmodifications, and conjugation with a non-peptidic extension (such aspolyethylene glycol, lipids, carbohydrates, nucleosides, nucleotides,nucleoside bases, various small molecules, or phosphate or sulfategroups). As used herein, the term “cyclic peptide mimetic” or “cyclicpolypeptide mimetic” refers to a peptide mimetic that has as part of itsstructure one or more cyclic features such as a loop, bridging moiety,and/or an internal linkage. As used herein, the term “bridging moiety”refers to a chemical moiety that chemically links one or a combinationof atoms on an amino acid to any other atoms outside of the amino acidresidue. For instance, in the case oa amino acid tertiary structure, abridging moiety may be a chemical moiety that chemicaly links one aminoacid side chain with another sequential or non-seqeuntial amino acidside chain.

In some embodiments, peptide or peptide mimetics are provided, whereinthe loop comprises a bridging moiety selected from the group consistingof:

wherein each X is independently N or CH, such that no ring contains morethan 2 N; each Z is independently a bond, NR, O, S, CH2, C(O)NR, NRC(O),S(O)vNR, NRS(O)v; each m is independently selected from 0, 1, 2, and 3;each vis independently selected from 1 and 2; each R is independentlyselected from Hand C₁-C₆; and each bridging moiety is connected to thepeptide by independently selected C₀-C₆ spacers.

In some embodiments, the PIF peptides of the disclosure are modified toproduce peptide mimetics by replacement of one or more naturallyoccurring side chains of the 20 genetically encoded amino acids (or Damino acids) with other side chains, for instance with groups such asalkyl, lower alkyl, cyclic 4-, 5-, 6-, to 7 membered alkyl, amide, amidelower alkyl, amide di (lower alkyl), lower alkoxy, hydroxy, carboxy andthe lower ester derivatives thereof, and with 4-, 5-, 6-, to 7 memberedheterocyclics. For example, proline analogs can be made in which thering size of the proline residue is changed from 5 members to 4, 6, or 7members. Cyclic groups can be saturated or unsaturated, and ifunsaturated, can be aromatic or nonaromatic. Heterocyclic groups cancontain one or more nitrogen, oxygen, and/or sulphur heteroatoms.Examples of such groups include the furazanyl,furyl, imidazolidinyl,imidazolyl, imidazolinyl, isothiazolyl, isoxazolyl, morpholinyl (e.g.morpholino), oxazolyl, piperazinyl (e.g. 1-piperazinyl), piperidyl (e.g.1-piperidyl, piperidino), pyranyl, pyrazinyl, pyrazolidinyl,pyrazolinyl, pyrazolyl, pyridazinyl, pyridyl, pyrimidinyl, pyrrolidinyl(e.g. 1-pyrrolidinyl), pyrrolinyl, pyrrolyl, thiadiazolyl, thiazolyl,thienyl, thiomorpholinyl (e.g. thiomorpholino), and triazolyl. Theseheterocyclic groups can be substituted or unsubstituted. Where a groupis substituted, the substituent can be alkyl, alkoxy, halogen, oxygen,or substituted or unsubstituted phenyl. Peptidomimetics may also haveamino acid residues that have been chemically modified byphosphorylation, sulfonation, biotinylation, or the addition or removalof other moieties.

In a further embodiment a compound of the formulaR₁-R₂-R₃-R₄-R₅-R₆-R₇-R₈-R₉-R₁₀-R₁₁-R₁₂-R₁₃-R₁₄-R₁₅, wherein R₁ is Met ora mimetic of Met, R₂ is Val or a mimetic of Val, R₃ is Arg or a mimeticof Arg, or any amino acid, R₄ is Ile or a mimetic of Ile, R₅ is Lys or amimetic of Lys, R₆ is Pro or a mimetic of Pro, R₇ is Gly or a mimetic ofGly, R₈ is Ser or a mimetic of Ser, R₉ is Ala or a mimetic of Ala, R₁₀is Asn or a mimetic of Asn, R₁₁ is Lys or a mimetic of Lys, R₁₂ is Proor a mimetic of Pro, R₁₃ is Ser or a mimetic of Ser, R₁₄ is Asp or amimetic of Asp and R₁₅ is Asp or a mimetic of Asp is provided. In afurther embodiment, a compound comprising the formulaR₁-R₂-R₃-R₄-R₅-R₆-R₇-R₈-R₉-R₁₀, wherein R₁ is Ser or a mimetic of Ser,R₂ is Gln or a mimetic of Gln, R₃ is Ala or a mimetic of Ala, R₄ is Valor a mimetic of Val, R₅ is Gln or a mimetic of Gln, R₆ is Glu or amimetic of Glu, R₇ is His or a mimetic of His, R₈ is Ala or a mimetic ofAla, R₉ is Ser or a mimetic of Ser, and R₁₀ is Thr or a mimetic of Thr;a compound comprising the formulaR₁-R₂-R₃-R₄-R₅-R₆-R₇-R₈-R₉-R₁₀-R₁₁-R₁₂-R₁₃-R₁₄-R₁₅-R₁₆-R₁₇-R₁₈, whereinR₁ is Ser or a mimetic of Ser, R₂ is Gly or a mimetic of Gly, R₃ is Ileor a mimetic of Ile, R₄ is Val or a mimetic of Val, R₅ is Ile or amimetic of Ile, R₆ is Tyr or a mimetic of Tyr, R₇ is Gln or a mimetic ofGln, R₈ is Tyr or a mimetic of Tyr, R₉ is Met or a mimetic of Met, R₁₀is Asp or a mimetic of Asp, R₁₁ is Asp or a mimetic of Asp, R₁₂ is Argor a mimetic of Arg, R₁₃ is Tyr or a mimetic of Tyr, R₁₄ is Val or amimetic of Val, R₁₅ is Gly or a mimetic of Gly, R₁₆ is Ser or a mimeticof Ser, R₁₇ is Asp or a mimetic of Asp and R₁₈ is Leu or a mimetic ofLeu; and a compound comprising the formula R₁-R₂-R₃-R₄-R₅-R₆-R₇-R₈-R₉,wherein R₁ is Val or a mimetic of Val, R₂ is Ile or a mimetic of Ile, R₃is Ile or a mimetic of Ile, R₄ is Ile or a mimetic of Ile, R₅ is Ala ora mimetic of Ala, R₆ is Gln or a mimetic of Gln, R₇ is Tyr or a mimeticof Tyr, R₈ is Met or a mimetic of Met, and R₉ is Asp or a mimetic of Aspis provided. In some embodiments, R₃ is not Arg or a mimetic of Arg.

A variety of techniques are available for constructing peptide mimeticswith the same or similar desired biological activity as thecorresponding native but with more favorable activity than the peptidewith respect to solubility, stability, and/or susceptibility tohydrolysis or proteolysis (see, e.g., Morgan & Gainor, Ann. Rep. Med.Chern. 24,243-252,1989). Certain peptidomimetic compounds are based uponthe amino acid sequence of the peptides of the disclosure. Often,peptidomimetic compounds are synthetic compounds having a threedimensional structure (i.e. a “peptide motif”) based upon thethree-dimensional structure of a selected peptide. The peptide motifprovides the peptidomimetic compound with the desired biologicalactivity, i.e., binding to PIF receptors, wherein the binding activityof the mimetic compound is not substantially reduced, and is often thesame as or greater than the activity of the native peptide on which themimetic is modeled. Peptidomimetic compounds can have additionalcharacteristics that enhance their therapeutic application, such asincreased cell permeability, greater affinity and/or avidity andprolonged biological half-life.

Peptidomimetic design strategies are readily available in the art (see,e.g., Ripka & Rich, Curr. Op. Chern. Bioi. 2,441-452,1998; Hruby et al.,Curr. Op.Chem. Bioi. 1,114-119,1997; Hruby & Baise, Curr. Med. Chern.9,945-970,2000). One class of peptidomimetics is a backbone that ispartially or completely non-peptide, but mimics the peptide backboneatom-for atom and comprises side groups that likewise mimic thefunctionality of the side groups of the native amino acid residues.Several types of chemical bonds, e.g., ester, thioester, thioamide,retroamide, reduced carbonyl, dimethylene and ketomethylene bonds, areknown in the art to be generally useful substitutes for peptide bonds inthe construction of protease-resistant peptidomimetics. Another class ofpeptidomimetics comprises a small non-peptide molecule that binds toanother peptide or protein, but which is not necessarily a structuralmimetic of the native peptide. Yet another class of peptidomimetics hasarisen from combinatorial chemistry and the generation of massivechemical libraries. These generally comprise novel templates which,though structurally unrelated to the native peptide, possess necessaryfunctional groups positioned on a nonpeptide scaffold to serve as“topographical” mimetics of the original peptide (Ripka & Rich, 1998,supra).

The first natural PIF compound identified, termed nPIF (SEQ ID NO: 1),is a 15 amino acid peptide. A synthetic version of this peptide, sPIF(SEQ ID NO:13), showed activity that was similar to the native peptide,nPIF (SEQ ID NO: I). This peptide is homologous to a small region of theCircumsporozoite protein, a malaria parasite. The second PIF peptide(SEQ ID NO:7), includes 13 amino acids and shares homology with a shortportion of a large protein named thyroid and retinoic acid transcriptionco-repressor, which is identified as a receptor-interacting factor,(SMRT); the synthetic version is sPIF-2 (SEQ ID NO:14). The thirddistinct peptide, nPIF-3 (SEQ ID NO:10), consists of 18 amino acids andmatches a small portion of reverse transcriptase; the synthetic versionof this peptide sPIF-3 is (SEQ ID NO:15). nPIF-4 (SEQ ID NO:12) shareshomology with a small portion of reverse transcriptase.

A list of PIF peptides, both natural and synthetic, are provided belowin Table 4. Antibodies to various PIF peptides and scrambled PIFpeptides are also provided.

TABLE 4 PIF Peptides (SEQ ID NO) Peptide Amino Acid SequenceSEQ ID NO: 1 nPIF-1₁₅ MVRIKPGSANKPSDD isolated native, matches region ofCircumsporozoite protein (Malaria) SEQ ID NO: 2 nPIF-1_((15-alter))MVRIKYGSYNNKPSD isolated native, matches region ofCircumsporozoite protein (Malaria) SEQ ID NO: 3 nPIF-1₍₁₃₎ MVRIKPGSANKPSisolated native, matches region of Circumsporozoite protein (Malaria)SEQ ID NO: 4 nPIF-1₍₉₎ MVRIKPGSA isolated native, matches region ofCircumsporozoite protein (Malaria) SEQ ID NO: 5 scrPIF-1₁₅GRVDPSNKSMPKDIA synthetic, scrambled amino acid sequence fromregion of Circumsporozoite protein Malaria SEQ ID NO: 6 nPIF-2₍₁₀₎SQAVQEHAST isolated native, matches region of humanretinoid and thyroid hormone receptor-SMRT SEQ ID NO: 7 nPIF-2₍₁₃₎SQAVQEHASTNMG isolated native, matches region of humanretinoid and thyroid hormone receptor (SMRT) SEQ ID NO: 8 scrPIF-2₍₁₃₎EVAQHSQASTMNG synthetic, scrambled amino acid sequence fromregion of human retinoid and thyroid hormone receptor SMRT SEQ ID NO: 9scrPIF-2₍₁₄₎ GQASSAQMNSTGVH SEQ ID NO: 10 nPIF-3₍₁₈₎ SGIVIYQYMDDRYVGSDLisolated native, matches region of Rev Trans SEQ ID NO: 11Neg control for GMRELQRSANKsynthetic, scrambled amino acid sequence from negPIF-1₍₁₅₎region of Circumsporozoite protein Malaria SEQ ID NO: 12 nP1F-4₍₉₎VIIIAQYMD isolated native, matches region of Rev Transantibody of native isolated nPIF-1₁₅ AbPIF-1₍₁₅₎ (SEQ ID NO: 13)sPIF-1₍₁₅₎ MVRIKPGSANKPSDD synthetic, amino acid sequence from region ofCircumsporozoite protein Malaria (SEQ ID NO: 14) sPIF-2₍₁₃₎SQAVQEHASTNMG synthetic, amino acid sequence from of humanretinoid and thyroid hormone receptor SMRT (SEQ ID NO: 15) sPIF-3₍₁₈₎SGIVIYQYMDDRYVGSDL synthetic, amino acid sequence from region ofCircumsporozoite protein Malaria (SEQ ID NO: 16) sPIF-1₍₉₎ MVRIKPGSAsynthetic, amino acid sequence from region ofCircumsporozoite protein Malaria antibody of native isolated nPIF-2₍₁₃₎AbPIF-2₍₁₃₎ antibody of native isolated nPIF-3₍₁₈₎ AbPIF-3₍₁₈₎(SEQ ID NO: 17) sPIF-4₍₉₎ VIIIAQYMD Synthetic SEQ ID NO: 18 sPIF-1₍₅₎MVRIK Synthetic SEQ ID NO: 19 sPIF-1₍₄₎ PGSA Synthetic SEQ ID NO: 20PIF (−3) MVXIKPGSANKPSDD SEQ ID NO: 21 PIF (−1) XVRIKPGSANKPSDDSEQ ID NO: 22 PIF (−1, −3) XVXIKPGSANKPSDD SEQ ID NO: 23 PIF (−6)MVRIKXGSANKPSDD SEQ ID NO: 24 PIF (−4) MVRXKPGSANKPSDD SEQ ID NO: 25PIF (−2) MXRIKPGSANKPSDD SEQ ID NO: 26 mut1 MVRIKEGSANKPSDDSEQ ID NO: 27 mut3 MVRGKPGSANKPSDD SEQ ID NO: 28 mut4 MERIKPGSANKPSDDSEQ ID NO: 29 mut5 AVRIKPGSANKPSDD n = native, s = synthetic, scr =scrambled, same AA, ( ) = number of AA, Ab = antibody, X = any aminoacid, except arginine

In some embodiments of the present disclosure, a PIF peptide isprovided. Such PIF peptides may be useful for acute radiation syndrome(ARS), delayed effects of acute radiation exposure, or conditionsrelated thereto.

In another embodiment, a pharmaceutical composition comprising a PIFpeptide is provided. In some embodiments, the pharmaceutical compositioncomprises a therapeutically effective amount of a PIF peptide or apharmaceutically acceptable salt thereof. In some embodiments, thepharmaceutical compsoitions is free of a peptide comprising any one ormore of the sequence identifiers of Table 4. In some embodiments, thepharmaceutical compsoitions is free of a peptide comprising orconsisting of SEQ ID NO:1.

In another embodiment, methods of treating acute radiation syndrome,delayed effects of acute radiation exposure, or conditions relatedthereto are provided. In a preferred embodiment, the method comprisesadministering an effective amount of a PIF peptide to a subject in needthereof.

In a further embodiment, a method for treating ARS comprisingadministering an effective amount of a PIF peptide in combination withone or more immunotherapeutic, anti-epileptic, diuretic, orantihypertensive drugs or compounds to a subject in need thereof isprovided. Such a combination may enhance the effectiveness of thetreatment of either component alone, or may provide less side effectsand/or enable a lower dose of either component.

PIF-1's action appears to be independent of TCR, calcium-channels or PKCpathways, mechanisms through which most immunosuppressive agents act,and CD4+/CD25+ cells (T reg) cells that are of relevance in variousautoimmune diseases. On the other hand, PIF-1's action may involveNFAT-1 suppression.

Ultimately, a novel embryo-derived peptide, PIF, creates a tolerogenicstate at low doses following short-term treatment leading to long-termprotection in several distinct severe autoimmune models. This effect isexerted without apparent toxicity.

For therapeutic treatment of the specified indications, a PIF peptidemay be administered as such, or can be compounded and formulated intopharmaceutical compositions in unit dosage form for parenteral,transdermal, rectal, nasal, local intravenous administration, or,preferably, oral administration. Such pharmaceutical compositions areprepared in a manner well known in the art and comprise at least oneactive PIF peptide associated with a pharmaceutically carrier. The term“active compound”, as used throughout this specification, refers to atleast one compound selected from compounds of the formulas orpharmaceutically acceptable salts thereof.

In such a composition, the active compound is known as the “activeingredient.” In making the compositions, the active ingredient willusually be mixed with a carrier, or diluted by a carrier, or enclosedwithin a carrier that may be in the form of a capsule, sachet, paper orother container. When the carrier serves as a diluent, it may be asolid, semisolid, or liquid material that acts as a vehicle, excipientof medium for the active ingredient. Thus, the composition can be in theform of tablets, pills, powders, lozenges, sachets, cachets, elixirs,emulsion, solutions, syrups, suspensions, soft and hard gelatincapsules, sterile injectable solutions, and sterile packaged powders.

The terms “pharmaceutical preparation” and “pharmaceutical composition”include preparations suitable for administration to mammals, e.g.,humans. When the compounds of the present disclosure are administered aspharmaceuticals to mammals, e.g., humans, they can be given per se or asa pharmaceutical composition containing, for example, from about 0.1 toabout 99.5% of active ingredient in combination with a pharmaceuticallyacceptable carrier.

The phrase “pharmaceutically acceptable” refers to molecular entitiesand compositions that are physiologically tolerable and do not typicallyproduce an allergic or similar untoward reaction, such as gastric upset,dizziness and the like, when administered to a human. Preferably, asused herein, the term “pharmaceutically acceptable” means approved by aregulatory agency of the Federal or a state government or listed in theU.S. Pharmacopeia or other generally recognized pharmacopeia for use inanimals, and more particularly in humans. In some embodiments, thepharmaceutical compsoitions comprising a PIF peptide, mimetic orpharmaceutically acceptable salt thereof and at least onepharmaceutically acceptable carrier.

The phrase “pharmaceutically acceptable carrier” is art recognized andincludes a pharmaceutically acceptable material, composition or vehicle,suitable for administering compounds of the present disclosure tomammals. The carriers include liquid or solid filler, diluent,excipient, solvent or encapsulating material, involved in carrying ortransporting the subject agent from one organ, or portion of the body,to another organ, or portion of the body. Each carrier must be“acceptable” in the sense of being compatible with the other ingredientsof the formulation and not injurious to the patient. Some examples ofmaterials which can serve as pharmaceutically acceptable carriersinclude: sugars, such as lactose, glucose and sucrose; starches, such ascorn starch and potato starch; cellulose, and its derivatives, such assodium carboxymethyl cellulose, ethyl cellulose and cellulose acetate;powdered tragacanth; malt; gelatin; talc; excipients, such as cocoabutter and suppository waxes; oils, such as peanut oil, cottonseed oil,safflower oil, sesame oil, olive oil, corn oil and soybean oil; glycols,such as propylene glycol; polyols, such as glycerin, sorbitol, mannitoland polyethylene glycol; esters, such as ethyl oleate and ethyl laurate;agar; buffering agents, such as magnesium hydroxide and aluminumhydroxide; alginic acid; pyrogen-free water; isotonic saline; Ringer'ssolution; ethyl alcohol; phosphate buffer solutions; and other non-toxiccompatible substances employed in pharmaceutical formulations. Suitablepharmaceutical carriers are described in “Remington's PharmaceuticalSciences” by E. W. Martin, which is incorporated herein by reference inits entirety. In some embodiments, the pharmaceutically acceptablecarrier is sterile and pyrogen-free water. In some embodiments, thepharmaceutically acceptable carrier is Ringer's Lactate, sometimes knownas lactated Ringer's solution.

Wetting agents, emulsifiers and lubricants, such as sodium laurylsulfate and magnesium stearate, as well as coloring agents, releaseagents, coating agents, sweetening, flavoring and perfuming agents,preservatives and antioxidants can also be present in the compositions.

Examples of pharmaceutically acceptable antioxidants include: watersoluble antioxidants, such as ascorbic acid, cysteine hydrochloride,sodium bisulfate, sodium metabisulfite, sodium sulfite and the like;oil-soluble antioxidants, such as ascorbyl palmitate, butylatedhydroxyanisole (BHA), butylated hydroxytoluene (BHT), lecithin, propylgallate, .alpha.-tocopherol, and the like; and metal chelating agents,such as citric acid, ethylenediamine tetraacetic acid (EDTA), sorbitol,tartaric acid, phosphoric acid, and the like.

Formulations of the present disclosure include those suitable for oral,nasal, topical, buccal, sublingual, rectal, vaginal and/or parenteraladministration. The formulations may conveniently be presented in unitdosage form and may be prepared by any methods well known in the art ofpharmacy. The amount of active ingredient that can be combined with acarrier material to produce a single dosage form will generally be thatamount of the compound that produces a therapeutic effect. Generally,out of one hundred percent, this amount will range from about 1 percentto about ninety-nine percent of active ingredient, preferably from about5 percent to about 70 percent, most preferably from about 10 percent toabout 30 percent.

Some examples of suitable carriers, excipients, and diluents includelactose, dextrose, sucrose, sorbitol, mannitol, starches, gum acacia,calcium phosphate alginates, calcium salicate, microcrystallinecellulose, polyvinylpyrrolidone, cellulose, tragacanth, gelatin, syrup,methyl cellulose, methyl- and propylhydroxybenzoates, tale, magnesiumstearate, water, and mineral oil. The formulations can additionallyinclude lubricating agents, wetting agents, emulsifying and suspendingagents, preserving agents, sweetening agents or flavoring agents. Thecompositions may be formulated so as to provide quick, sustained, ordelayed release of the active ingredient after administration to thepatient by employing procedures well known in the art.

For oral administration, a compound can be admixed with carriers anddiluents, molded into tablets, or enclosed in gelatin capsules. Themixtures can alternatively be dissolved in liquids such as 10% aqueousglucose solution, isotonic saline, sterile water, or the like, andadministered intravenously or by injection.

The local delivery of inhibitory amounts of active compound for thetreatment of immune disorders can be by a variety of techniques thatadminister the compound at or near the targeted site. Examples of localdelivery techniques are not intended to be limiting but to beillustrative of the techniques available. Examples include localdelivery catheters, site specific carriers, implants, direct injection,or direct applications, such as topical application.

Local delivery by an implant describes the surgical placement of amatrix that contains the pharmaceutical agent into the affected site.The implanted matrix releases the pharmaceutical agent by diffusion,chemical reaction, or solvent activators.

For example, in some aspects, the disclosure is directed to apharmaceutical composition comprising a PIF peptide, and apharmaceutically acceptable carrier or diluent, or an effective amountof pharmaceutical composition comprising a PIF peptide.

Specific modes of administration will depend on the indication. Theselection of the specific route of administration and the dose regimenis to be adjusted or titrated by the clinician according to methodsknown to the clinician in order to obtain the optimal clinical response.The amount of compound to be administered is that amount which istherapeutically effective. The dosage to be administered will depend onthe characteristics of the subject being treated, e.g., the particularmammal or human treated, age, weight, health, types of concurrenttreatment, if any, and frequency of treatments, and can be easilydetermined by one of skill in the art (e.g., by the clinician).

Pharmaceutical formulations containing the compounds of the presentdisclosure and a suitable carrier can be solid dosage forms whichinclude, but are not limited to, tablets, capsules, cachets, pellets,pills, powders and granules; topical dosage forms which include, but arenot limited to, solutions, powders, fluid emulsions, fluid suspensions,semi-solids, ointments, pastes, creams, gels,jellies, and foams; andparenteral dosage forms which include, but are not limited to,solutions, suspensions, emulsions, and dry powder; comprising aneffective amount of a polymer or copolymer of the present disclosure. Itis also known in the art that the active ingredients can be contained insuch formulations with pharmaceutically acceptable diluents, fillers,disintegrants, binders, lubricants, surfactants, hydrophobic vehicles,water soluble vehicles, emulsifiers, buffers, humectants, moisturizers,solubilizers, preservatives and the like. The means and methods foradministration are known in the art and an artisan can refer to variouspharmacologic references for guidance. For example, ModernPharmaceutics, Banker & Rhodes, Marcel Dekker, Inc. (1979); and Goodman& Gilman's The Pharmaceutical Basis of Therapeutics, 6th Edition,MacMillan Publishing Co., New York (1980) can be consulted.

The compounds of the present disclosure can be formulated for parenteraladministration by injection, e.g., by bolus injection or continuousinfusion. The compounds can be administered by continuous infusionsubcutaneously over a predetermined period of time. Formulations forinjection can be presented in unit dosage form, e.g., in ampoules or inmulti-dose containers, with an added preservative. The compositions cantake such forms as suspensions, solutions or emulsions in oily oraqueous vehicles, and can contain formulatory agents such as suspending,stabilizing and/or dispersing agents.

For oral administration, the compounds can be formulated readily bycombining these compounds with pharmaceutically acceptable carriers wellknown in the art. Such carriers enable the compounds of the disclosureto be formulated as tablets, pills, dragees, capsules, liquids, gels,syrups, slurries, suspensions and the like, for oral ingestion by apatient to be treated. Pharmaceutical preparations for oral use can beobtained by adding a solid excipient, optionally grinding the resultingmixture, and processing the mixture of granules, alter adding suitableauxiliaries, if desired, to obtain tablets or dragee cores. Suitableexcipients include, but are not limited to, fillers such as sugars,including, but not limited to, lactose, sucrose, mannitol, and sorbitol;cellulose preparations such as, but not limited to, maize starch, wheatstarch, rice starch, potato starch, gelatin, gum tragecanth, methylcellulose, hydroxypropylmethyl-celllose, sodium carboxymethylcellulose,and polyvinylpyrrolidone (PVP). If desired, disintegrating agents can beadded, such as, but not limited to, the cross-linked polyvinylpyrrolidone, agar, or alginic acid or a salt thereof such as sodiumalginate.

Dragee cores can be provided with suitable coatings. For this purpose,concentrated sugar solutions can be used, which can optionally containgum arabic, talc, polyvinyl pyrrolidone, carbopol gel, polyethyleneglycol, and/or titanium dioxide, lacquer solutions, and suitable organicsolvents or solvent mixtures. Dyestuffs or pigments can be added to thetablets or dragee coatings for identification or to characterizedifferent combinations of active compound doses.

Pharmaceutical preparations which can be used orally include, but arenot limited to, push-fit capsules made of gelatin, as well as soft,scaled capsules made of gelatin and a plasticizer, such as glycerol orsorbitol. The push-fit capsules can contain the active ingredients inadmixture with filler such as, e.g., lactose, binders such as, e.g.,starches, and/or lubricants such as, e.g., talc or magnesium stearateand, optionally, stabilizers. In soft capsules, the active compounds canbe dissolved or suspended in suitable liquids, such as fatty oils,liquid paraffin, or liquid polyethylene glycols. In addition,stabilizers can be added. All formulations for oral administrationshould be in dosages suitable for such administration.

For buccal administration, the compositions can take the form of, e.g.,tablets or lozenges formulated in a conventional manner.

For administration by inhalation, the compounds for use according to thepresent disclosure are conveniently delivered in the form of an aerosolspray presentation from pressurized packs or a nebulizer, with the useof a suitable propellant, e.g., dichlorodifluoromethane,trichlorofluoromethane, dichlorotetrafluoroethane, carbon dioxide orother suitable gas. In the case of a pressurized aerosol the dosage unitcan be determined by providing a valve to deliver a metered amount.Capsules and cartridges of, e.g., gelatin for use in an inhaler orinsufflator can be formulated containing a powder mix of the compoundand a suitable powder base such as lactose or starch.

The compounds of the present disclosure can also be formulated in rectalcompositions such as suppositories or retention enemas, e.g., containingconventional suppository bases such as cocoa butter or other glycerides.

In addition to the formulations described previously, the compounds ofthe present disclosure can also be formulated as a depot preparation.Such long acting formulations can be administered by implantation (forexample subcutaneously or intramuscularly) or by intramuscularinjection.

Depot injections can be administered at about 1 to about 6 months orlonger intervals. Thus, for example, the compounds can be formulatedwith suitable polymeric or hydrophobic materials (for example as anemulsion in an acceptable oil) or ion exchange resins, or as sparinglysoluble derivatives, for example, as a sparingly soluble salt.

In transdermal administration, the compounds of the present disclosure,for example, can be applied to a plaster, or can be applied bytransdermal, therapeutic systems that are consequently supplied to theorganism.

Pharmaceutical compositions of the compounds also can comprise suitablesolid or gel phase carriers or excipients. Examples of such carriers orexcipients include but are not limited to calcium carbonate, calciumphosphate, various sugars, starches, cellulose derivates, gelatin, andpolymers such as, e.g., polyethylene glycols.

For parenteral administration, analog can be, for example, formulated asa solution, suspension, emulsion or lyophilized powder in associationwith a pharmaceutically acceptable parenteral vehicle. Examples of suchvehicles are water, saline, Ringer's solution, dextrose solution, and 5%human serum albumin. Liposomes and nonaqueous vehicles such as fixedoils may also be used. The vehicle or lyophilized powder may containadditives that maintain isotonicity (e.g., sodium chloride, mannitol)and chemical stability (e.g., buffers and preservatives). Theformulation is sterilized by commonly used techniques. For example, aparenteral composition suitable for administration by injection isprepared by dissolving 1.5% by weight of analog in 0.9% sodium chloridesolution.

The present disclosure relates to routes of administration includeintramuscular, sublingual, intravenous, intraperitoneal, intrathecal,intravaginal, intraurethral, intradermal, intrabuccal, via inhalation,via nebulizer and via subcutaneous injection. Alternatively, thepharmaceutical composition may be introduced by various means into cellsthat are removed from the individual. Such means include, for example,microprojectile bombardment and liposome or other nanoparticle device.

Solid dosage forms for oral administration include capsules, tablets,pills, powders and granules. In solid dosage forms, the analogs aregenerally admixed with at least one inert pharmaceutically acceptablecarrier such as sucrose, lactose, starch, or other generally regarded assafe (GRAS) additives. Such dosage forms can also comprise, as is normalpractice, an additional substance other than an inert diluent, e.g.,lubricating agent such as magnesium state. With capsules, tablets, andpills, the dosage forms may also comprise a buffering agent. Tablets andpills can additionally be prepared with enteric coatings, or in acontrolled release form, using techniques know in the art.

Liquid dosage forms for oral administration include pharmaceuticallyacceptable emulsions, solutions, suspensions and syrups, with theelixirs containing an inert diluent commonly used in the art, such aswater. These compositions can also include one or more adjuvants, suchas wetting agent, an emulsifying agent, a suspending agent, a sweeteningagent, a flavoring agent or a perfuming agent.

In another embodiment of the invention the composition of the inventionis used to treat a patient suffering from, or susceptible to Type Iadult or juvenile diabetes, multiple sclerosis, Crohn's, or autoimmunehepatitis.

One of skill in the art will recognize that the appropriate dosage ofthe compositions and pharmaceutical compositions may vary depending onthe individual being treated and the purpose. For example, the age, bodyweight, and medical history of the individual patient may affect thetherapeutic efficacy of the therapy. Further, a lower dosage of thecomposition may be needed to produce a transient cessation of symptoms,while a larger dose may be needed to produce a complete cessation ofsymptoms associated with the disease, disorder, or indication. Acompetent physician can consider these factors and adjust the dosingregimen to ensure the dose is achieving the desired therapeutic outcomewithout undue experimentation. It is also noted that the clinicianand/or treating physician will know how and when to interrupt, adjust,and/or terminate therapy in conjunction with individual patientresponse. Dosages may also depend on the strength of the particularanalog chosen for the pharmaceutical composition.

The dose of the composition or pharmaceutical compositions may vary. Thedose of the composition may be once per day. In some embodiments,multiple doses may be administered to the subject per day. In someembodiments, the total dosage is administered in at least twoapplication periods. In some embodiments, the period can be an hour, aday, a month, a year, a week, or a two-week period. In an additionalembodiment of the invention, the total dosage is administered in two ormore separate application periods, or separate doses over the course ofan hour, a day, a month, a year, a week, or a two-week period.

In some embodiments, subjects can be administered the composition inwhich the composition is provided in a daily dose range of about 0.0001mg/kg to about 5000 mg/kg of the weight of the subject. The doseadministered to the subject can also be measured in terms of totalamount of PIF peptide or PIF analog or pharmaceutically acceptable saltthereof administered per day. In some embodiments, a subject isadministered from about 0.001 to about 3000 milligrams of PIF peptide orPIF analog or pharmaceutically acceptable salt thereof per day. In someembodiments, a subject is administered up to about 2000 milligrams ofPIF peptide or PIF analog or pharmaceutically acceptable salt thereofper day. In some embodiments, a subject is administered up to about 1800milligrams of PIF peptide or PIF analog or pharmaceutically acceptablesalt thereof per day. In some embodiments, a subject is administered upto about 1600 milligrams of PIF peptide or PIF analog orpharmaceutically acceptable salt thereof per day. In some embodiments, asubject is administered up to about 1400 milligrams of PIF peptide orPIF analog or pharmaceutically acceptable salt thereof per day. In someembodiments, a subject is administered up to about 1200 milligrams ofPIF peptide or PIF analog or pharmaceutically acceptable salt thereofper day. In some embodiments, a subject is administered up to about 1000milligrams of PIF peptide or PIF analog or pharmaceutically acceptablesalt thereof per day. In some embodiments, a subject is administered upto about 800 milligrams of PIF peptide or PIF analog or pharmaceuticallyacceptable salt thereof per day. In some embodiments, a subject isadministered from about 0.001 milligrams to about 700 milligrams of PIFpeptide or PIF analog or pharmaceutically acceptable salt thereof perdose. In some embodiments, a subject is administered up to about 700milligrams of PIF peptide or PIF analog per dose. In some embodiments, asubject is administered up to about 600 milligrams of PIF peptide or PIFanalog or pharmaceutically acceptable salt thereof per dose. In someembodiments, a subject is administered up to about 500 milligrams of PIFpeptide or PIF analog or pharmaceutically acceptable salt thereof perdose. In some embodiments, a subject is administered up to about 400milligrams of PIF peptide or PIF analog or pharmaceutically acceptablesalt thereof per dose. In some embodiments, a subject is administered upto about 300 milligrams of PIF peptide or PIF analog or pharmaceuticallyacceptable salt thereof per dose. In some embodiments, a subject isadministered up to about 200 milligrams of PIF peptide or PIF analog orpharmaceutically acceptable salt thereof per dose. In some embodiments,a subject is administered up to about 100 milligrams of PIF peptide orPIF analog or pharmaceutically acceptable salt thereof per dose. In someembodiments, a subject is administered up to about 50 milligrams of PIFpeptide or PIF analog or pharmaceutically acceptable salt thereof perdose.

In some embodiments, subjects can be administered the composition inwhich the composition comprising a PIF peptide or PIF analog orpharmaceutically acceptable salt thereof is administered in a daily doserange of about 0.0001 mg/kg to about 5000 mg/kg of the weight of thesubject. In some embodiments, the composition comprising a PIF analog orpharmaceutically acceptable salt thereof is administered in a dailydosage of up to about 450 mg/kg of the weight of the subject. In someembodiments, the composition comprising a PIF peptide or PIF analog orpharmaceutically acceptable salt thereof is administered in a dailydosage of up to about 400 mg/kg of the weight of the subject. In someembodiments, the composition comprising a PIF peptide or PIF analog orpharmaceutically acceptable salt thereof is administered in a dailydosage of up to about 350 mg/kg of the weight of the subject. In someembodiments, the composition comprising a PIF peptide or PIF analog orpharmaceutically acceptable salt thereof is administered in a dailydosage of up to about 300 mg/kg of the weight of the subject. In someembodiments, the composition comprising a PIF peptide or PIF analog orpharmaceutically acceptable salt thereof is administered in a dailydosage of up to about 250 mg/kg of the weight of the subject. In someembodiments, the composition comprising PIF peptide or a PIF analog orpharmaceutically acceptable salt thereof is administered in a dailydosage of up to about 200 mg/kg of the weight of the subject. In someembodiments, the composition comprising PIF peptide or a PIF analog orpharmaceutically acceptable salt thereof is administered in a dailydosage of up to about 150 mg/kg of the weight of the subject. In someembodiments, the composition comprising a PIF peptide or a PIF analog orpharmaceutically acceptable salt thereof is administered in a dailydosage of up to about 100 mg/kg of the weight of the subject. In someembodiments, the composition comprising a PIF peptide or a PIF analog orpharmaceutically acceptable salt thereof is administered in a dailydosage of up to about 50 mg/kg of the weight of the subject. In someembodiments, the composition comprising PIF peptide or a PIF analog orpharmaceutically acceptable salt thereof is administered in a dailydosage of up to about 25 mg/kg of the weight of the subject.

In some embodiments, the composition comprising a PIF peptide or a PIFanalog or pharmaceutically acceptable salt thereof is administered in adaily dosage of up to about 10 mg/kg of the weight of the subject. Insome embodiments, the composition comprising PIF peptide or a PIF analogor pharmaceutically acceptable salt thereof is administered in a dailydosage of up to about 5 mg/kg of the weight of the subject. In someembodiments, the composition comprising PIF peptide or a PIF analog orpharmaceutically acceptable salt thereof is administered in a dailydosage of up to about 1 mg/kg of the weight of the subject. In someembodiments, the composition comprising a PIF peptide or a PIF analog orpharmaceutically acceptable salt thereof is administered in a dailydosage of up to about 0.1 mg/kg of the weight of the subject.

In some embodiments, the composition comprising a PIF analog orpharmaceutically acceptable salt thereof is administered in a dailydosage of up to about 0.01 mg/kg of the weight of the subject. In someembodiments, the composition comprising a PIF analog or pharmaceuticallyacceptable salt thereof is administered in a daily dosage of up to about0.001 mg/kg of the weight of the subject. The dose administered to thesubject can also be measured in terms of total amount of a PIF peptideor PIF analog administered per day.

In some embodiments, a subject in need thereof is administered fromabout 1 ng to about 500 μg of analog or pharmaceutically salt thereofper day. In some embodiments, a subject in need thereof is administeredfrom about 1 ng to about 10 ng of analog or pharmaceutically saltthereof per day. In some embodiments, a subject in need thereof isadministered from about 10 ng to about 20 ng of analog orpharmaceutically salt thereof per day. In some embodiments, a subject inneed thereof is administered from about 10 ng to about 100 ng of analogor pharmaceutically salt thereof per day. In some embodiments, a subjectin need thereof is administered from about 100 ng to about 200 ng ofanalog or pharmaceutically salt thereof per day. In some embodiments, asubject in need thereof is administered from about 200 ng to about 300ng of analog or pharmaceutically salt thereof per day. In someembodiments, a subject in need thereof is administered from about 300 ngto about 400 ng of analog or pharmaceutically salt thereof per day. Insome embodiments, a subject in need thereof is administered from about400 ng to about 500 ng of analog or pharmaceutically salt thereof perday. In some embodiments, a subject in need thereof is administered fromabout 500 ng to about 600 ng of analog or pharmaceutically salt thereofper day. In some embodiments, a subject in need thereof is administeredfrom about 600 ng to about 700 ng of analog or pharmaceutically saltthereof per day. In some embodiments, a subject in need thereof isadministered from about 800 ng to about 900 ng of analog orpharmaceutically salt thereof per day. In some embodiments, a subject inneed thereof is administered from about 900 ng to about 1 μg of analogor pharmaceutically salt thereof per day. In some embodiments, a subjectin need thereof is administered from about 1 μg to about 100 μg ofanalog or pharmaceutically salt thereof per day. In some embodiments, asubject in need thereof is administered from about 100 μg to about 200of analog or pharmaceutically salt thereof per day. In some embodiments,a subject in need thereof is administered from about 200 μg to about 300μg of analog or pharmaceutically salt thereof per day. In someembodiments, a subject in need thereof is administered from about 300 toabout 400 μg of analog or pharmaceutically salt thereof per day. In someembodiments, a subject in need thereof is administered from about 400 μgto about 500 μg of analog or pharmaceutically salt thereof per day. Insome embodiments, a subject in need thereof is administered from about500 μg to about 600 μg of analog or pharmaceutically salt thereof perday. In some embodiments, a subject in need thereof is administered fromabout 600 μg to about 700 μg of analog or pharmaceutically salt thereofper day. In some embodiments, a subject in need thereof is administeredfrom about 800 μg to about 900 μg of analog or pharmaceutically saltthereof per day. In some embodiments, a subject in need thereof isadministered from about 900 μg to about 1 mg of analog orpharmaceutically salt thereof per day.

In some embodiments, a subject in need thereof is administered fromabout 0.0001 to about 3000 milligrams of a PIF peptide or PIF analog orpharmaceutically salt thereof per day. In some embodiments, a subject isadministered up to about 2000 milligrams of a PIF peptide or PIF analogor pharmaceutically salt thereof day. In some embodiments, a subject isadministered up to about 1800 milligrams of a PIF peptide or PIF analogor pharmaceutically salt thereof per day. In some embodiments, a subjectis administered up to about 1600 milligrams of a PIF peptide or PIFanalog or pharmaceutically salt thereof per day. In some embodiments, asubject is administered up to about 1400 milligrams of a PIF peptide orPIF analog or pharmaceutically salt thereof per day. In someembodiments, a subject is administered up to about 1200 milligrams of aPIF peptide or PIF analog or pharmaceutically salt thereof per day. Insome embodiments, a subject is administered up to about 1000 milligramsof a PIF peptide or PIF analog or pharmaceutically salt thereof per day.In some embodiments, a subject is administered up to about 800milligrams of a PIF peptide or PIF analog or pharmaceutically saltthereof per day. In some embodiments, a subject is administered fromabout 0.0001 milligrams to about 700 milligrams of a PIF peptide or PIFanalog or pharmaceutically salt thereof per dose. In some embodiments, asubject is administered up to about 700 milligrams of a PIF peptide orPIF analog or pharmaceutically salt thereof per dose. In someembodiments, a subject is administered up to about 600 milligrams of aPIF peptide or PIF analog or pharmaceutically salt thereof per dose. Insome embodiments, a subject is administered up to about 500 milligramsof a PIF peptide or PIF analog or pharmaceutically salt thereof perdose. In some embodiments, a subject is administered up to about 400milligrams of a PIF peptide or PIF analog or pharmaceutically saltthereof per dose. In some embodiments, a subject is administered up toabout 300 milligrams of a PIF peptide or PIF analog or pharmaceuticallysalt thereof per dose. In some embodiments, a subject is administered upto about 200 milligrams of a PIF peptide or PIF analog orpharmaceutically salt thereof per dose. In some embodiments, a subjectis administered up to about 100 milligrams of a PIF peptide or PIFanalog or pharmaceutically salt thereof per dose. In some embodiments, asubject is administered up to about 50 milligrams of a PIF peptide orPIF analog or pharmaceutically salt thereof per dose. In someembodiments, a subject is administered up to about 25 milligrams of aPIF peptide or PIF analog or pharmaceutically salt thereof per dose. Insome embodiments, a subject is administered up to about 15 milligrams ofa PIF peptide or PIF analog or pharmaceutically salt thereof per dose.

In some embodiments, a subject is administered up to about 10 milligramsof a PIF peptide or PIF analog or pharmaceutically salt thereof perdose. In some embodiments, a subject is administered up to about 5milligrams of a PIF peptide or PIF analog or pharmaceutically saltthereof per dose. In some embodiments, a subject is administered up toabout 1 milligram of a PIF peptide or PIF analog or pharmaceuticallysalt thereof per dose. In some embodiments, a subject is administered upto about 0.1 milligrams of a PIF peptide or PIF analog orpharmaceutically salt thereof per dose. In some embodiments, a subjectis administered up to about 0.001 milligrams of a PIF peptide or PIFanalog or pharmaceutically salt thereof per dose.

The dose administered to the subject can also be measured in terms oftotal amount of a PIF peptide or PIF analog or pharmaceutically saltthereof administered per ounce of liquid prepared. In some embodiments,the PIF peptide or PIF analog or pharmaceutically salt thereof is at aconcentration of about 2.5 grams per ounce of solution. In someembodiments, the PIF peptide or PIF analog or pharmaceutically saltthereof is at a concentration of about 2.25 grams per ounce of solution.In some embodiments, the PIF peptide or PIF analog or pharmaceuticallysalt thereof is at a concentration of about 2.25 grams per ounce ofsolution. In some embodiments, the PIF peptide or PIF analog orpharmaceutically salt thereof is at a concentration of about 2.0 gramsper ounce of solution. In some embodiments, the PIF peptide or PIFanalog or pharmaceutically salt thereof is at a concentration of about1.9 grams per ounce of solution. In some embodiments, the PIF peptide orPIF analog or pharmaceutically salt thereof is at a concentration ofabout 1.8 grams per ounce of solution. In some embodiments, the PIFanalog or pharmaceutically salt thereof is at a concentration of about1.7 grams per ounce of solution. In some embodiments, the PIF peptide orPIF analog or pharmaceutically salt thereof is at a concentration ofabout 1.6 grams per ounce of solution. In some embodiments, the PIFpeptide or PIF analog or pharmaceutically salt thereof is at aconcentration of about 1.5 grams per ounce of solution. In someembodiments, the PIF peptide or PIF analog or pharmaceutically saltthereof is at a concentration of about 1.4 grams per ounce of solution.In some embodiments, the PIF peptide or PIF analog or pharmaceuticallysalt thereof is at a concentration of about 1.3 grams per ounce ofsolution. In some embodiments, the PIF peptide or PIF analog orpharmaceutically salt thereof is at a concentration of about 1.2 gramsper ounce of solution. In some embodiments, the PIF peptide or PIFanalog or pharmaceutically salt thereof is at a concentration of about1.1 grams per ounce of solution. In some embodiments, the PIF peptide orPIF analog or pharmaceutically salt thereof is at a concentration ofabout 1.0 grams per ounce of solution.

In some embodiments, the PIF peptide or PIF analog or pharmaceuticallysalt thereof is at a concentration of about 0.9 grams per ounce ofsolution. In some embodiments, the PIF peptide or PIF analog orpharmaceutically salt thereof is at a concentration of about 0.8 gramsper ounce of solution. In some embodiments, the PIF peptide or PIFanalog or pharmaceutically salt thereof is at a concentration of about0.7 grams per ounce of solution. In some embodiments, the PIF peptide orPIF analog or pharmaceutically salt thereof is at a concentration ofabout 0.6 grams per ounce of solution. In some embodiments, the PIFpeptide or PIF analog or pharmaceutically salt thereof is at aconcentration of about 0.5 grams per ounce of solution. In someembodiments, the PIF peptide or PIF analog or pharmaceutically saltthereof is at a concentration of about 0.4 grams per ounce of solution.In some embodiments, the PIF peptide or PIF analog or pharmaceuticallysalt thereof is at a concentration of about 0.3 grams per ounce ofsolution. In some embodiments, the PIF peptide or PIF analog orpharmaceutically salt thereof is at a concentration of about 0.2 gramsper ounce of solution. In some embodiments, the PIF peptide or PIFanalog or pharmaceutically salt thereof is at a concentration of about0.1 grams per ounce of solution. In some embodiments, the PIF peptide orPIF analog or pharmaceutically salt thereof is at a concentration ofabout 0.01 grams per ounce of solution. In some embodiments, the PIFpeptide or PIF analog or pharmaceutically salt thereof is at aconcentration of about 0.001 grams per ounce of solution prepared. Insome embodiments, the PIF peptide or PIF analog or pharmaceutically saltthereof is at a concentration of about 0.0001 grams per ounce ofsolution prepared. In some embodiments, the PIF peptide or PIF analog orpharmaceutically salt thereof is at a concentration of about 0.00001grams per ounce of solution prepared. In some embodiments, the PIFpeptide or PIF analog or pharmaceutically salt thereof is at aconcentration of about 0.000001 grams per ounce of solution prepared.

Dosage may be measured in terms of mass amount of analog per liter ofliquid formulation prepared. One skilled in the art can increase ordecrease the concentration of the analog in the dose depending upon thestrength of biological activity desired to treat or prevent anyabove-mentioned disorders associated with the treatment of subjects inneed thereof. For instance, some embodiments of the invention caninclude up to 0.00001 grams of analog per 5 mL of liquid formulation andup to about 10 grams of analog per 5 mL of liquid formulation.

In some embodiments the pharmaceutical compositions of the claimedinvention comprises at least one or a plurality of active agents otherthan the PIF peptide, analog of pharmaceutically acceptable saltthereof. In some embodiments the active agent is covalently linked tothe PIF peptide or PIF analog disclosed herein optionally by a proteasecleavable linker (including by not limited to Pro-Pro or Cituline-Valinedi-α-amino acid linkers). In some embodiments, the one or plurality ofactive agents is one or a combination of compounds chosen from: ananti-inflammatory compound, alpha-adrenergic agonist, antiarrhythmiccompound, analgesic compound, and an anesthetic compound.

TABLE 5 Examples of anti-inflammatory compounds include: aspirincelecoxib diclofenac diflunisal etodolac ibuprofen indomethacinketoprofen ketorolac nabumetone naproxen oxaprozin piroxicam salsalatesulindac tolmetin Examples of alpha-adrenergic agonists include:Methoxamine Methylnorepinephrine Midodrine Oxymetazoline MetaraminolPhenylephrine Clonidine (mixed alpha2-adrenergic and imidazoline-I1receptor agonist) Guanfacine, (preference for alpha2A-subtype ofadrenoceptor) Guanabenz (most selective agonist for alpha2-adrenergic asopposed to imidazoline-I1) Guanoxabenz (metabolite of guanabenz)Guanethidine (peripheral alpha2-receptor agonist) Xylazine, TizanidineMedetomidine Methyldopa Fadolmidine Dexmedetomidine Examples ofantiarrhythmic compounds include: Amiodarone (Cordarone, Pacerone)Bepridil Hydrochloride (Vascor) Disopyramide (Norpace) Dofetilide(Tikosyn) Dronedarone (Multaq) Flecainide (Tambocor) Ibutilide (Corvert)Lidocaine (Xylocaine) Procainamide (Procan, Procanbid) Propafenone(Rythmol) Propranolol (Inderal) Quinidine (many trade names) Sotalol(Betapace) Tocainide (Tonocarid) Examples of analgesic compound include:codeine hydrocodone (Zohydro ER), oxycodone (OxyContin, Roxicodone),methadone hydromorphone (Dilaudid, Exalgo), morphine (Avinza, Kadian,MSIR, MS Contin), and fentanyl (Actiq, Duragesic) Examples of anestheticcompounds include: Desflurane Isoflurane Nitrous oxide Sevoflurane Xenon

The compounds of the present disclosure can also be administered incombination with other active ingredients, such as, for example,adjuvants, or other compatible drugs or compounds where such combinationis seen to be desirable or advantageous in achieving the desired effectsof the methods described herein. When exposing the PIF peptide to anycells, tissue, or organ prior to transplantation, exposure may beanywhere from about 1 to about 12 hours. In some embodiments, the stepof exposing PIF to pre-condition cells prior to transplant is from about2 to about 4 hours. In some embodiments, the step of exposing PIF topre-condition cells prior to transplant is about 1, 2, 3, 4, 5, 6, 7, 8,9, 10, 11, or 12 or more hours. In some embodiments, the step ofexposing PIF to the organ, tissue, or cells prior to transplant occursany where from about 1 to about 48 hours before transplant. In someembodiments, the step of exposing PIF to the organ, tissue, or cellsprior to transplant occurs is about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11,12, 15, 20, 25, 35, 45, or about 50 hours before transplant occurs. Insome embodiments, the PIF peptide is exposed to the organ, tissue orcells for a time and under conditions sufficient to increase theviability of the organ, tissue or cells, increase the likelihood ofsuccessful transplantation, reduce recipient acceptance. In someembodiments, the organ, tissue or cells is exposed to one or acombination of pharmaceutical compositions disclosed herein for no lessthan 1, 2, 3, 4, 6, 7, 8, 10 hours and/or at about room temperature. Insome embodiments, the organ, tissue or cells is exposed to one or acombination of pharmaceutical compositions disclosed herein for no lessthan 1, 2, 3, 4, 6, 7, 8, 10 hours and/or at about 4 degree Celsius. Insome embodiments, the organ, tissue or cells is exposed to one or acombination of pharmaceutical compositions disclosed herein at fromabout 4 degree Celsius to about 40 degrees Celsius.

As used herein, the terms “acute radiation syndrome” and “acuteradiation sickness” may be used interchangeably to refer to any acute orchronic symptom associated with lethal or sub-lethal exposure toradiation.

As used herein, the term “islet cells” refers to cells from anyhormone-producing region of the pancreas.

As used herein, the term “hematopoietic cells” refers to any cell thatmay give rise to any type of blood cell within an organism. In someembodiments, the hematopoietic cell is a hematopoietic stem cell. Insome embodiments, the hematopoietic cell is a pluripotent precursor cellcapable of being differentiated into a blood cell. In some embodiments,the hematopoietic cell is a bone marrow cell. In some embodiments, thehematopoietic cell is a blood cell that was differentiated from aninduced pluripotent stem cell, an embryonic cell, or a mesenchymal cell.

As used herein, the term “adrenal cells” refers to any cell from anyhormone-producing region of the adrenal gland.

As used herein, the term “heart cells” refers to any cell from anyportion of the heart or blood vessels. In some embodiments, thehematopoietic cell is a heart cell that was differentiated from aninduced pluripotent stem cell, an embryonic stem cell, or a mesenchymalstem cell.

As used herein, the term “pre-condition” refers to the process oftreating an organ, tissue, or cell prior to its transplantation or use.Any organ, tissue, or cell may be pre-conditioned one or more times.

As used herein, the term “adrenal cell disorder” refers to anydysfunction of any adrenal cell, and/or symptoms associated with suchdysfunction. In some embodiments, the adrenal cell disorder is Cushing'sdisease requiring an adrenal cell transplant. In some embodiments, thedisorder is any disease requiring an adrenal cell transplant.

As used herein, the term “blood disorder” refers to any dysfunction ofany blood or blood component, and/or symptoms associated with suchdysfunction. In some embodiments, the blood disorder is a blood cancer,such as leukemia. In some embodiments, the subject may have or suspectedof having an immune disorder such as multiple scelorsis or Crohn'sdisease such that transplantation of the bone marrow or circulatingimmune cell population may treat or prevent progression of the disease.

As used herein, the term “heart disorder” refers to any dysfunction ofany heart or heart component, and/or symptoms associated with suchdysfunction. In some embodiments, the heart disorder is congestive heartfailure.

Embodiments of the invention are directed to the use of PIF to treatacute radiation syndrome (ARS). In certain embodiments, methods oftreating ARS comprise administering a PIF peptide to a subject, tissue,organ, or cells in need thereof. In some embodiments, ARS may be fromintentional exposure to radiation, for example, in the case of radiationtherapy for the treatment of cancer. In some embodiments, PIF may beadministered before, after or in conjunction with radiation therapy or acombination thereof. In some embodiments, ARS may be from unintentionalexposure to radiation. In some embodiments, a method of treating ARSfollowing exposure to radiation may comprise administering PIF to asubject tissue, organ, or cells in need thereof. In some embodiments, amethod of treating radiation-induced organ damage may compriseadministering PIF to a subject tissue, organ, or cells in need thereof.In some embodiments, the organ damage may be to any organ in the body.In some embodiments, the organ damage may be to a vital organ of thebody. In some embodiments, the organ damage may be damage to the skin,brain, heart, lungs, kidneys, spleen, liver, gastrointestinal tract, orpancreas. In some embodiments, a method of treating delayed effects ofacute radiation exposure (DEARE) may comprise administering PIF to asubject, tissue, organ, or cells in need thereof. In some embodiments, amethod of inhibiting or preventing the development of ARS after exposureto radiation may comprise administering PIF to a subject tissue, organ,or cells in need thereof. In some embodiments, a method of treating ARSmay comprise transplanting bone marrow into a subject in need thereof,wherein the bone marrow is exposed to PIF prior to transplantation. Insome embodiments, the subject does not receive an organ, tissue or celltransplant. In some embodiments, the subject does not receive a bonemarrow transplant.

Some embodiments are directed to the use of PIF as an immune modulatoryagent for the improved acceptance of bone marrow or any other organ,tissue, or cell in autologous transplants, allogeneic transplants,semi-allogeneic transplants, or xenotransplants. An improved acceptanceof bone marrow or any other organ, tissue, or cell may allow an enhancedsurvival of transplant patients and a wider use of autologoustransplantation, allogeneic transplantation, semi-allogeneictransplantation, and xenotransplantation. Some embodiments are directedto a method of increasing engraftment of the transplanted organ, tissue,or cells comprising administering a PIF peptide to a subject in needthereof. Some embodiments are directed to a method of increasingengraftment of the transplanted organ, tissue, or cells comprisingpre-exposing the organ, tissue, or cells to the PIF peptide prior totransplantation. In some embodiments, the subject may be in need of theorgan, tissue, or cell transplantation to treat ARS.

In some embodiments, a method of treating ARS in subjects with cancerfollowing exposure to radiation comprises administering PIF to thesubject. In some embodiments, the subject's exposure to radiation mayhave been intentional, such as when radiation therapy is used for thetreatment of cancer. In some embodiments, the subject with cancer willnot receive a transplant of bone marrow or any other organ before,during, or following the administration of PIF.

In some embodiments, a method of preventing elevation in expression ofchemokines and cytokines after exposure to radiation comprisesadministering PIF to a subject in need thereof. In some embodiments, amethod of improving immune cell function following exposure to radiationcomprises administering PIF to a subject in need thereof. In someembodiments, PIF may be self-administered. In some embodiments, a methodof increasing hepatic function following exposure to radiation comprisesadministering PIF to a subject in need thereof. In some embodiments, amethod of normalizing hepatic enzyme levels following exposure toradiation comprises administering PIF to a subject in need thereof. Insome embodiments, a method of treating or preventing a stochastic effectof radiation may comprise administering PIF to a subject in needthereof. In some embodiments, stochastic effects of radiation mayinclude cancer, tumors, genetic damage or a combination thereof.

In some embodiments, PIF may be administered as a preventative, as aconcomitant, shortly after exposure, after organ damage, or acombination thereof. In such embodiments, PIF may be an effectiveARS/DEARE countermeasure covering the spectrum of damage from caseswhere there is no pre-event information, and extending to those caseswhere belated information may become available and damage has occurred.In some embodiments, PIF may not only address the immune aspects such asthose present in ARS but also the related pathological aspects thatoccur in general due to such exposure to radiation (organ damage) andeven at later time post-transplant when PIF was shown to provideprotection against graft vs host (GVHD) as well as graft vs. leukemia(GVL)—frequent delayed effects of acute radiation exposure (DEARE). Inother instances, in case of a hoax or non-specific information, wherePIF may be administered as a preventative, minimal or no adversereaction may be expected, as it is believed that PIF is inherentlynontoxic.

In the foregoing embodiments, exposure to radiation may include exposureto lethal doses of radiation. In the foregoing embodiments, exposure toradiation may include exposure to non-lethal doses of radiation. In theforegoing embodiments, the radiation dose may be from about 100 rads toabout 3000 rads, from about 200 rads to about 3000 rads, from about 500rads to about 3000 rads, from about 800 rads to about 3000 rads, fromabout 1000 rads to about 3000 rads, from about 100 rads to about 1000rads, from about 200 rads to about 1000 rads, from about 500 rads toabout 1000 rads, from about 100 rads to about 5000 rads or from about100 rads to about 6000 rads, and any range between any of these values,including endpoints.

In the foregoing embodiments, the PIF peptide may be administered priorto, concurrently with or following exposure to radiation.

In some embodiments, the PIF peptide is administered or is pre-exposedin a therapeutically effective amount. In some embodiments, the PIFpeptide is administered after the subject undergoes transplant, beforethe subject undergoes transplant, while the subject undergoestransplant, or a combination thereof. In some embodiments, the subjectmay receive secondary treatment, which may include secondaryadministration of a PIF peptide, before the subject undergoestransplant, while the subject undergoes transplant, or a combinationthereof.

In the foregoing embodiments, the PIF peptide may be administered at adose of about 0.01 mg/kg/day, about 0.1 mg/kg/day, about 0.5 mg/kg/day,0.75 mg/kg/day, 1 mg/kg/day, 2 mg/kg/day, 4 mg/kg/day, 6 mg/kg/day, 8mg/kg/day, 10 mg/kg/day, 20 mg/kg/day, or any range between any of thesevalues, including endpoints. Such doses may be administered as a singledose or as divided doses in a single day.

In the foregoing embodiments, the PIF may be administered once, for alimited period of time or as a maintenance therapy (over an extendedperiod of time until the condition is ameliorated, cured or for the lifeof the subject). A limited period of time may be for 1 week, 2 weeks, 3weeks, 4 weeks and up to one year, including any period of time betweensuch values, including endpoints. In some embodiments, the PIF peptidemay be administered for about 1 day, for about 3 days, for about 1 week,for about 10 days, for about 2 weeks, for about 18 days, for about 3weeks, or for any range between any of these values, including endpoints

In the foregoing embodiments, the PIF may be administered once daily,twice daily, three times daily, four times daily or more.

In the foregoing embodiments, the PIF peptide may be administered beforeexposure to radiation, within about 6 hours of exposure to radiation,within about 12 hours of exposure to radiation, within about 18 hours ofexposure to radiation, within about 24 hours of exposure to radiation,within about 30 hours of exposure to radiation, within about 36 hours ofexposure to radiation, within about 42 hours of exposure to radiation,within about 48 hours of exposure to radiation, or within any rangebetween any of these values, including endpoints.

In some embodiments, the PIF is administered or provided as apharmaceutical composition comprising a PIF peptide, as defined above,and a pharmaceutically acceptable carrier or diluent, or an effectiveamount of a pharmaceutical composition comprising a compound as definedabove.

The methods disclosed herein can be used with any of the compounds,compositions, preparations, and kits disclosed herein.

In some embodiments, the disclosure relates to methods for treatingacute radiation syndrome comprising administering an effective amount ofthe compositions described herein to a subject in need thereof.

In some embodiments, the disclosure relates to methods for treatingacute radiation syndrome following radiation exposure comprisingtransplanting bone marrow to a subject in need thereof, wherein the bonemarrow is pre-exposed to an effective amount of the compositionsdescribed herein.

In some embodiments, the disclosure relates to methods for increasingengraftment of a transplanted organ, tissue, or cells comprisingtransplanting the organ, tissue, or cell into a subject in need thereof,wherein the organ, tissue, or cell is pre-exposed to an effective amountof the compositions described herein prior to transplantation.

In some embodiments, the disclosure relates to a method of increasingthe likelihood of acceptance of a transplant of a donor organ, tissue,or cell into a subject, comprising exposing the organ, tissue, or cellto one or more compositions described herein prior to transplanting theorgan, tissue, or cell into the subject.

In some embodiments, the disclosure relates to a method of reducing thelikelihood of rejection of an engrafted tissue, comprising exposing thetissue to one or more of the compositions described herein prior totransplanting the tissue into a subject.

In some embodiments, the disclosure relates to a method of increasingproduction of hematopoietic cells in a subject having a depleted numberof hematopoietic cells, comprising administering one or morepharmaceutical compositions described herein.

In an embodiment, the composition is administered once a day to asubject in need thereof. In another embodiment, the composition isadministered every other day, every third day or once a week. In anotherembodiment, the composition is administered twice a day. In stillanother embodiment, the composition is administered three times a day orfour times a day. In a further embodiment, the composition isadministered at least once a day for at least 1, 2, 3, 4, 5, 6, 7, 8, 9,10, 11 or 12 weeks. In still a further embodiment, the composition isadministered at least once a day for a longer term such as at least 4,6, 8, 10, 12 or 24 months. Administration in some embodiments includesbut is not limited to a dosage of 10-50 mg of composition at a frequencyof minimum 1, 2, 3 or 4 times per day. In some embodiments, thecompositions is administered once a week, once every other week or oncea month. Optionally, administration continues until all symptoms areresolved and cleared by medical personnel.

In some embodiments, the composition is administered within 1, 2, 3, 5or 7 days of exposure to radiation. In other embodiments, thecomposition is administered within 1, 2, 3, 5 or 7 days of theappearance of symptoms of ARS.

In some embodiments, the composition is administered at least once a dayuntil the condition has ameliorated to where further treatment is notnecessary. In another embodiment, the composition is administered untilall symptoms of the condition are resolved. In further embodiments, thecomposition is administered for at least 1, 2, 3, 6, 8, 10 or 12 or 24months after the subject is asymptomatic.

The compositions of the present disclosure are useful and effective whenadministered to treat acute radiation syndrome, as well as topre-condition organs, cells, or tissues prior to transplantation. Theamount of each component present in the composition will be the amountthat is therapeutically effective, i.e., an amount that will result inthe effective treatment of the condition (e.g., ARS) when administered.The therapeutically effective amount will vary depending on the subjectand the severity and nature of the injury and can be determinedroutinely by one of ordinary skill in the art.

In some embodiments, the disclosure relates to a method of treating orpreventing any of the indications set forth in U.S. Pat. Nos. 7,723,289,7,723,290, 8,222,211, 8,454,967, 9,097,725, (each of which areincorporated by reference in their entireties) comprising administeringcompositions or pharmaceutical compositions comprising any one orplurality of PIF peptides, analogs, or pharmaceutically acceptable saltsthereof disclosed herein.

In some methods, the disclosure relates to a method of stimulating thedifferentiation and/or proliferation of stem cells in a subject in needthereof comprising administering compositions or pharmaceuticalcompositions comprising any one or plurality of PIF peptides, analogs,or pharmaceutically acceptable salts thereof disclosed herein.

In some embodiments, the disclosure relates to any of the methodsdisclosed in U.S. Pat. Nos. 7,273,708, 7,695,977, 7,670,852, 7,670,851,7,678,582, 7,670,850, 8,012,700 (each of which are incorporated byreference in their entireties) comprising administering compositions orpharmaceutical compositions comprising any one or plurality of PIFpeptides, analogs, or pharmaceutically acceptable salts thereofdisclosed herein. This disclosure also incorporates by reference intheir entireties U.S. Pat. Nos. 7,789,289, 7,723,290, 8,222,211, and8,454,967.

In some embodiments, the disclosure relates to a pharmaceuticalcomposition comprising a therapeutically effective amount or dose of atleast one PIF peptide, an analog thereof, or a pharmaceuticallyacceptable salt thereof, and a pharmaceutically acceptable carrier forthe treatment of acute radiation syndrome.

In some embodiments, the disclosure relates to the use of atherapeutically effective amount or dose of any one or plurality ofcompositions disclosed herein comprising at least one PIF peptide, ananalog thereof, or a pharmaceutically acceptable salt thereof, and apharmaceutically acceptable carrier in the manufacture of a medicamentfor the pre-condition of organs, tissues, or cells prior totransplantation.

In some embodiments, the disclosure relates to the use of apharmaceutical composition comprising a therapeutically effective amountor dose at least one PIF peptide, an analog thereof, or apharmaceutically acceptable salt thereof, and a pharmaceuticallyacceptable carrier in the manufacture of a medicament for the treatmentacute radiation syndrome.

In some embodiments, the disclosure relates to a method of inducing animmunomodulation effect in a subject in need thereof, when subject hasbeen or is suspect of having acute radiation syndrome.

In some embodiments, the disclosure relates to a method of treatingacute radiation syndrome by administering at least one or a plurality ofcompositions disclosed herein comprising PIF peptide, an analog thereof,or a pharmaceutically acceptable salt thereof.

In some embodiments, the disclosure relates to a method of treatingacute radiation syndrome by administering a therapeutically effectiveamount or dose of one or a plurality of compositions disclosed hereincomprising at least one PIF peptide, an analog thereof, or apharmaceutically acceptable salt thereof.

In some embodiments, the disclosure relates to a method of treatingacute radiation syndrome by administration of a pharmaceuticalcomposition comprising a therapeutically effective amount or dose of atleast one PIF peptide, an analog thereof, or a pharmaceuticallyacceptable salt thereof, and a pharmaceutically acceptable carrier.

In some embodiments, the disclosure relates to a pharmaceuticalcomposition comprising a therapeutically effective amount or dose of atleast one PIF peptide, an analog thereof, or a pharmaceuticallyacceptable salt thereof, and a pharmaceutically acceptable carrier forthe treatment of acute radiation syndrome.

In some embodiments, the disclosure relates to the use of atherapeutically effective amount or dose of any one or plurality ofcompositions disclosed herein comprising at least one PIF peptide, ananalog thereof, or a pharmaceutically acceptable salt thereof, and apharmaceutically acceptable carrier in the manufacture of a medicamentfor the treatment of acute radiation syndrome.

In some embodiments, the disclosure relates to the use of apharmaceutical composition comprising a therapeutically effective amountor dose at least one PIF peptide, an analog thereof, or apharmaceutically acceptable salt thereof, and a pharmaceuticallyacceptable carrier in the manufacture of a medicament for the treatmentof acute radiation syndrome.

In some embodiments, the disclosure relates to a method of inducing animmunomodulation effect in a subject in need thereof, when subject hasor is suspected of having acute radiation syndrome.

In some embodiments, the disclosure relates to a method of treatinggraft-versus-host disease by administering at least one or a pluralityof compositions disclosed herein comprising PIF peptide, an analogthereof, or a pharmaceutically acceptable salt thereof.

In some embodiments, the disclosure relates to a method of treatinggraft-versus-host disease by administering a therapeutically effectiveamount or dose of one or a plurality of compositions disclosed hereincomprising at least one PIF peptide, an analog thereof, or apharmaceutically acceptable salt thereof.

In some embodiments, the disclosure relates to a method of treatinggraft-versus-host disease by administration of a pharmaceuticalcomposition comprising a therapeutically effective amount or dose of atleast one PIF peptide, an analog thereof, or a pharmaceuticallyacceptable salt thereof, and a pharmaceutically acceptable carrier.

In some embodiments, the disclosure relates to a pharmaceuticalcomposition comprising a therapeutically effective amount or dose of atleast one PIF peptide, an analog thereof, or a pharmaceuticallyacceptable salt thereof, and a pharmaceutically acceptable carrier forthe treatment of graft-versus-host disease.

In some embodiments, the disclosure relates to the use of atherapeutically effective amount or dose of any one or plurality ofcompositions disclosed herein comprising at least one PIF peptide, ananalog thereof, or a pharmaceutically acceptable salt thereof, and apharmaceutically acceptable carrier in the manufacture of a medicamentfor the treatment of graft-versus-host disease.

In some embodiments, the disclosure relates to the use of apharmaceutical composition comprising a therapeutically effective amountor dose at least one PIF peptide, an analog thereof, or apharmaceutically acceptable salt thereof, and a pharmaceuticallyacceptable carrier in the manufacture of a medicament for the treatmentof graft-versus-host disease.

In some embodiments, the disclosure relates to a method of inducing animmunomodulation effect in a subject in need thereof, when subject hasor is suspected of having graft-versus-host disease.

In some embodiments, the disclosure relates to a method of treatinginflammation by administering at least one or a plurality ofcompositions disclosed herein comprising PIF peptide, an analog thereof,or a pharmaceutically acceptable salt thereof.

In some embodiments, the disclosure relates to a method of treatinginflammation by administering a therapeutically effective amount or doseof one or a plurality of compositions disclosed herein comprising atleast one PIF peptide, an analog thereof, or a pharmaceuticallyacceptable salt thereof.

In some embodiments, the disclosure relates to a method of treatinginflammation by administration of a pharmaceutical compositioncomprising a therapeutically effective amount or dose of at least onePIF peptide, an analog thereof, or a pharmaceutically acceptable saltthereof, and a pharmaceutically acceptable carrier.

In some embodiments, the disclosure relates to a pharmaceuticalcomposition comprising a therapeutically effective amount or dose of atleast one PIF peptide, an analog or mimetic thereof, or apharmaceutically acceptable salt thereof, and a pharmaceuticallyacceptable carrier for the treatment of inflammation.

In some embodiments, the disclosure relates to the use of atherapeutically effective amount or dose of any one or plurality ofcompositions disclosed herein comprising at least one PIF peptide, ananalog thereof, or a pharmaceutically acceptable salt thereof, and apharmaceutically acceptable carrier in the manufacture of a medicamentfor the treatment of inflammation.

In some embodiments, the disclosure relates to the use of apharmaceutical composition comprising a therapeutically effective amountor dose at least one PIF peptide, an analog thereof, or apharmaceutically acceptable salt thereof, and a pharmaceuticallyacceptable carrier in the manufacture of a medicament for the treatmentof inflammation.

In some embodiments, the disclosure relates to a method of inducing animmunomodulation effect in a subject in need thereof, when subject hasbeen or is suspect of having inflammation.

In some embodiments, the disclosure relates to a method of treatingauto-immune disease by administering at least one or a plurality ofcompositions disclosed herein comprising PIF peptide, an analog thereof,or a pharmaceutically acceptable salt thereof.

In some embodiments, the disclosure relates to a method of treatingauto-immune disease by administering a therapeutically effective amountor dose of one or a plurality of compositions disclosed hereincomprising at least one PIF peptide, an analog thereof, or apharmaceutically acceptable salt thereof.

In some embodiments, the disclosure relates to a method of treatingauto-immune disease by administration of a pharmaceutical compositioncomprising a therapeutically effective amount or dose of at least onePIF peptide, an analog thereof, or a pharmaceutically acceptable saltthereof, and a pharmaceutically acceptable carrier.

In some embodiments, the disclosure relates to a pharmaceuticalcomposition comprising a therapeutically effective amount or dose of atleast one PIF peptide, an analog thereof, or a pharmaceuticallyacceptable salt thereof, and a pharmaceutically acceptable carrier forthe treatment of auto-immune disease.

In some embodiments, the disclosure relates to the use of atherapeutically effective amount or dose of any one or plurality ofcompositions disclosed herein comprising at least one PIF peptide, ananalog thereof, or a pharmaceutically acceptable salt thereof, and apharmaceutically acceptable carrier in the manufacture of a medicamentfor the treatment of auto-immune disease.

In some embodiments, the disclosure relates to the use of apharmaceutical composition comprising a therapeutically effective amountor dose at least one PIF peptide, an analog thereof, or apharmaceutically acceptable salt thereof, and a pharmaceuticallyacceptable carrier in the manufacture of a medicament for the treatmentof auto-immune disease.

In some embodiments, the disclosure relates to a method of inducing animmunomodulation effect in a subject in need thereof, when subject hasbeen or is suspect of having auto-immune disease.

In some embodiments, the disclosure relates to a method of treatinginflammation disorders by administering at least one or a plurality ofcompositions disclosed herein comprising PIF peptide, an analog thereof,or a pharmaceutically acceptable salt thereof.

In some embodiments, the disclosure relates to a method of treatinginflammation disorders by administering a therapeutically effectiveamount or dose of one or a plurality of compositions disclosed hereincomprising at least one PIF peptide, an analog thereof, or apharmaceutically acceptable salt thereof.

In some embodiments, the disclosure relates to a method of treatinginflammation disorders by administration of a pharmaceutical compositioncomprising a therapeutically effective amount or dose of at least onePIF peptide, an analog thereof, or a pharmaceutically acceptable saltthereof, and a pharmaceutically acceptable carrier.

In some embodiments, the disclosure relates to a pharmaceuticalcomposition comprising a therapeutically effective amount or dose of atleast one PIF peptide, an analog thereof, or a pharmaceuticallyacceptable salt thereof, and a pharmaceutically acceptable carrier forthe treatment of inflammation disorders.

In some embodiments, the disclosure relates to the use of atherapeutically effective amount or dose of any one or plurality ofcompositions disclosed herein comprising at least one PIF peptide, ananalog thereof, or a pharmaceutically acceptable salt thereof, and apharmaceutically acceptable carrier in the manufacture of a medicamentfor the treatment of inflammation disorders.

In some embodiments, the disclosure relates to the use of apharmaceutical composition comprising a therapeutically effective amountor dose at least one PIF peptide, an analog thereof, or apharmaceutically acceptable salt thereof, and a pharmaceuticallyacceptable carrier in the manufacture of a medicament for the treatmentof inflammation disorders.

In some embodiments, the disclosure relates to a method of inducing animmunomodulation effect in a subject in need thereof, when subject hasbeen or is suspect of having inflammation disorders.

In some embodiments, the disclosure relates to a method of treatingrepetitive strain injuries by administering at least one or a pluralityof compositions disclosed herein comprising PIF peptide, an analogthereof, or a pharmaceutically acceptable salt thereof.

According to some embodiments of the invention, the formulation may besupplied as part of a kit. In some embodiments, the kit comprisescomprising a PIF peptide and/or a PIF analog or pharmaceuticallyacceptable salt thereof, the PIF peptide and/or a PIF analog orpharmaceutically acceptable salt thereof comprises a non-natural aminoacid or is at least 70% homologous to SEQ ID NO:20. In anotherembodiment, the kit comprises a pharmaceutically acceptable salt of ananalog with a rehydration mixture. In another embodiment, thepharmaceutically acceptable salt of an analog are in one container whilethe rehydration mixture is in a second container. The rehydrationmixture may be supplied in dry form, to which water or other liquidsolvent may be added to form a suspension or solution prior toadministration. Rehydration mixtures are mixtures designed to solubilizea lyophilized, insoluble salt of the invention prior to administrationof the composition to a subject takes at least one dose of a purgative.In another embodiment, the kit comprises a pharmaceutically acceptablesalt in orally available pill form.

The kit may contain two or more containers, packs, or dispenserstogether with instructions for preparation and administration. In someembodiments, the kit comprises at least one container comprising thepharmaceutical composition or compositions described herein and a secondcontainer comprising a means for delivery of the compositions such as asyringe . In some embodiments, the kit comprises a compositioncomprising an analog in solution or lyophilized or dried and accompaniedby a rehydration mixture. In some embodiments, the analog andrehydration mixture may be in one or more additional containers.

The compositions included in the kit may be supplied in containers ofany sort such that the shelf-life of the different components arepreserved, and are not adsorbed or altered by the materials of thecontainer. For example, suitable containers include simple bottles thatmay be fabricated from glass, organic polymers, such as polycarbonate,polystyrene, polypropylene, polyethylene, ceramic, metal or any othermaterial typically employed to hold reagents or food; envelopes, thatmay consist of foil-lined interiors, such as aluminum or an alloy. Othercontainers include test tubes, vials, flasks, and syringes. Thecontainers may have two compartments that are separated by a readilyremovable membrane that upon removal permits the components of thecompositions to mix. Removable membranes may be glass, plastic, rubber,or other inert material.

Kits may also be supplied with instructional materials. Instructions maybe printed on paper or other substrates, and/or may be supplied as anelectronic-readable medium, such as a floppy disc, CD-ROM, DVD-ROM, zipdisc, videotape, audio tape, or other readable memory storage device.Detailed instructions may not be physically associated with the kit;instead, a user may be directed to an internet web site specified by themanufacturer or distributor of the kit, or supplied as electronic mail.

In another embodiment, a packaged kit is provided that contains thepharmaceutical formulation to be administered, i.e., a pharmaceuticalformulation containing PIF peptide and/or a PIF analog orpharmaceutically acceptable salt thereof, a container (e.g., a vial, abottle, a pouch, an envelope, a can, a tube, an atomizer, an aerosolcan, etc.), optionally sealed, for housing the formulation duringstorage and prior to use, and instructions for carrying out drugadministration in a manner effective to treat any one or more of theindications disclosed herein. The instructions will typically be writteninstructions on a package insert, a label, and/or on other components ofthe kit.

Depending on the type of formulation and the intended mode ofadministration, the kit may also include a device for administering theformulation (e.g., a transdermal delivery device). The administrationdevice may be a dropper, a swab, a stick, or the nozzle or outlet of anatomizer or aerosol can. The formulation may be any suitable formulationas described herein. For example, the formulation may be an oral dosageform containing a unit dosage of the active agent, or a gel or ointmentcontained within a tube. The kit may contain multiple formulations ofdifferent dosages of the same agent. The kit may also contain multipleformulations of different active agents.

The present kits will also typically include means for packaging theindividual kit components, i.e., the pharmaceutical dosage forms, theadministration device (if included), and the written instructions foruse. Such packaging means may take the form of a cardboard or paper box,a plastic or foil pouch, etc.

This disclosure and embodiments illustrating the method and materialsused may be further understood by reference to the followingnon-limiting examples.

EXAMPLES Materials and Methods:

The following materials and methods were used to conduct the experimentsdescribed herein.

ARS murine models: C57BL/6 mice (6-7 or 8-9 week old females) and F1(C57BL/6×Balb/c) mice (10-11 week old females) were obtained from HarlanLaboratories Ltd (Israel). The study was conducted under ethicalconditions approved by the Institutional Animal Welfare Committee of theHebrew University of Jerusalem. Mice were kept and monitored inpathogen-free conditions. Mice from the C57BL/6 strain underwentwhole-body irradiation by a single dose of sub-lethal (6Gry) or lethal(10Gry) whole-body irradiation at a dose rate (0.3Gry/min), using aclinical 6MEV (Linear Varian CL-6), Varian medical systems, Palo Alto,Calif., USA.

sPIF*: Synthetic PIF (MVRIKPGSANKPSDD) was obtained from BiosynthesisLewisville, N.J. USA. The peptide was purified to >95% as documented byHPLC/mass spectrometry (sPIF proprietary).

Statistical analysis: Data from in vivo studies are represented asmean±SEM. Data from in vitro studies are represented as mean±SD. Singlecomparisons to control were made using two-tailed Student's t-test orMann-Whitney test. One-way repeated measures ANOVA followed byBonferroni's Multiple Comparison Test were used for multigroup design.P<0.05 was considered to be statistically significant. Data handling andstatistical processing was performed using Microsoft Excel and GraphPadPrism Software. Gene expression was determined based on the AACt methodand calculated by the qBASE+ software. Results are expressed as foldchange from a standard reference sample included in each run. Theanalyses of gene expression were calculated via a non-parametricMann-Whitney U Test. P<0.05 was considered significant. Colon globalgene analysis was carried out using heat map followed by individualgenes determining differences among the groups setting P<0.05 assignificant.

Long-term sub-lethal sPIF experiments in ARS: Analysis of hematopoieticrecovery: Mice underwent whole-body irradiation (6Gry). After 24 hrs PIFor PBS (1 mg/kg/day) was administered continuously (0.25 ml/hr) toC57BL/6 mice for two weeks using subcutaneously implanted Alzet osmoticpumps (Model 1002, Durect Corp., CA). This was followed by two weeks ofobservation post-therapy without any added therapy. No antibiotics wereadministered in any of the experiments. The hematopoietic profile ofeach mouse was examined weekly until sacrifice. Weekly up to four weeksat sacrifice, mice were tail-bled and 100 μl blood was collected intoEDTA coated capillary tubes. A CBC with differential was performed usinga validated BC-2800 Vet Auto Hematology Analyzer (Mindray, Mahwah, N.J.USA). Results were compared with normal mice.

Short-term sub-lethal sPIF experiments in ARS: hematopoiesis colon andsystemic cytokines: Mice were irradiated (6 or 7 Gry). 24 or 48 hrspost-irradiation 0.75 mg/kg, sPIF was injected subcutaneously twicedaily for three days. At the end of the experiment mice were sacrificedand colon tissue removed for histology, Illumina global genome analysis,and RT-PCR. In addition, the serum was removed for cytokine analysis.Results were compared with normal mice. For sub-lethal long-term ARSexperiments, sPIF (1 mg/kg/day: Alzet osmotic pump; Model 1002, DurectCorp., CA) or PBS was injected for 14 days followed by 14 days ofobservation post-therapy. Hematopoietic profiles were examined weekly bycollecting 100 μl tail blood. CBC with differential was performed usinga validated BC-2800 Vet Auto Hematology Analyzer (Mindray, Mahwah, N.J.USA).

sPIF induced long-term hematopoietic recovery post-lethal irradiationand semi-allogeneic BMT: CBC and tibia bone histology analysis: Micewere irradiated (10 Gry), lethal dose. One day post-irradiation, bonemarrow (BM) mononuclear cells from donor mice F1 (C57BL/6×Balb/c) werecollected by flushing the femur and tibia bone with PBS (BiologicalIndustries). Mononuclear cells were isolated by using Lymphoprep method.A total of 8*106 BM cells were administered to the tail vein ofirradiated recipient mice. Following BMT, mice were monitored daily forloss of weight, ruffled skin, and survival as previously described. Oncea week mice up to four weeks at sacrifice were tail-bled and 100 μlblood was collected into EDTA coated capillary tubes. CBC withdifferential was performed using a validated BC-2800 Vet Auto HematologyAnalyzer (Mindray, Mahwah, N.J. USA). Results were compared with normalmice.

sPIF preconditioned allogeneic BMT: Whether BM preconditioned with PIFcan improve BMT engraftment without further therapy after BMT wasdetermined. Mice underwent (10Gry) whole body irradiation. After 24 hrs,BM cells were exposed to sPIF for only 2 hrs before transplantation,followed by washing off the cells prior to inoculation to the tail veinof recipient mice. As control BM cells were exposed to PBS beforetransplant. After transplantation, mice were followed without anyfurther therapy for up to 4 weeks. Following sacrifice total WBC countand lymphocytes concentrations were carried out. Results were comparedto PBS—control.

sPIF effects MSC regulatory function: CFSE stained murine splenocyteswere activated with anti-CD3 antibodies, were cultured for four days (ina 50:1 ratio) and were exposed to MSCs previously incubated (2 h) withsPIF or control. Cell proliferation was analyzed using Flow Cytometry.Data was analyzed the % proliferating cells by comparing the sPIFpre-treated MSCs as compared to control (activated splenocytes withoutMSCs).

sPIF-treated femur bone histological analysis: Mice were irradiated10Gry and after 24 hrs sPIF treatment was initiated, lasting for 14 days(Alzet pump) following by 14 days post-therapy. sPIF-preconditioned BMTwithout further therapy was studied as well. At the end of the study WBCand femur bone samples were obtained from mice following sacrifice andfixed in 4% neutral-buffered formalin. Samples were decalcified thenembedded in paraffin, cut into 10-micron thick sections, and stainedwith hematoxylin and eosin (H&E). Results were compared with PBS treatedmice and with normal mice.

sPIF-treated serum cytokine evaluation: Circulating cytokine levels fromperipheral blood were determined by using Mouse Th1/Th2 10plexFlowCytomix Multiplex kit (eBioscience, SanDiego, Calif., USA) accordingto the manufacturer's protocol.

sPIF-treated colon histology and crypt depth determination: Followingsub-lethal irradiation (6Gry) sPIF treatment started at 24 or 48 hrspost-irradiation lasting 2 or 3. Following sacrifice, the colon washarvested and washed extensively in PBS to remove intestinal contents.The jejunum was fixed in 10% neutral buffered formalin prior to paraffinembedding. Samples were processed into 5 mm sections for hematoxylin andeosin (Fisher Scientific, Pittsburgh, Pa.) routinely and crypt depth wasdetermined. Notably, crypt depth was reported as a marker of recoveryafter radiation exposure. For this analysis a BX51 microscope (Olympus,Tokyo, Japan) equipped with a digital camera was used and imagesacquired using a 10× objective. The images were analyzed using ImageJsoftware as previously reported. The effect of sPIF treatment after 24and 48 hrs post-therapy was compared with PBS and normal mice.

sPIF gene analysis: RT-qPCR analysis macrophages and colon tissue: sPIFtargets macrophages and is effective in GVHD model reducing oxidativestress genes in the liver. Therefore, gene expression in colon samplesfrom sub-lethal short-term mice after 24-48 post-radiation (6Gry) wasdetermined following exposure to sPIF compared to PBS and normal mice.This was carried out in two independent sets of experiments. Inaddition, a number of genes were determined also in macrophagesfollowing exposure for 24 h to sPIF in vitro determining macrophagepolarity. Total RNA was extracted using RNeasy® Mini Kit columns(QIAGEN, Hilden, Germany) according to the manufacturer's protocols. 1μg of total RNA was used to synthesize cDNA using High-Capacity cDNA kit(Applied Biosystems, Gran Island, N.Y., USA) according to manufacturer'sinstructions as reported previously. Detection of transcript levels ofB7H1, NOS2, and Arg-1, was performed using the TaqMan Gene ExpressionAssay Kit (Applied (Applied Biosystems). HPRT-1 was used as ahousekeeping gene transcript to normalized endogenous control. Allprimers were purchased from Applied Biosystems. Real-Time PCR reactionwas carried out using the ABI Prism 7900 Sequence system (AppliedBiosystems). Data was analyzed by StepOne Software version 2.2 (AppliedBiosystems). DataAssist Software v3.01. Data sets p-values were adjustedusing Benjamin-Hochberg method.

sPIF protective signaling pathway: Colon Illumina gene Array: Toround-up elucidating sPIF's protective signaling pathways, a global genearray was performed. Following sub-lethal radiation (6Gry), at 24 hrssPIF injection twice daily for 72 hrs were administered. The effect ofsPIF following radiation was compared with PBS-treated control. Normalmice without irradiation served as an additional control. Followingsacrifice, 30 mg of colon tissue (N=4-5 treatment group) was excised andhomogenized in a Fastprep 120 tissue homogenizer (30 s at 4.0 m/sec) incell lysis buffer (Qiagen, Hombrechtikon, Switzerland). Total RNAs wereextracted from cells using PureLink RNA Mini Kit (Ambion, catalog number12183018A). Total RNA (250 ng) was amplified into cRNA using TotalPrepRNA amplification kit (AMIL1791, Ambion) following manufacture'sinstruction. After amplification, 1.5 μg of cRNA was mixed with thehybridization controls and it was hybridized to MouseRef-8 array(BD-202-0202, Illumina, USA). The array was hybridized for 16 hrs in ahybridization oven with a rocking platform at 58° C. The array chip thenwent through a series of washes before it was stained withstreptavidin-Cy3. After the staining, it went through a final wash anddrying. The array was scanned using the Illumina HiScan Scanner.

sPIF-induced macrophage shift polarization: Cell isolation and in vitromacrophage differentiation: To determine the ability of sPIF to inducemacrophage polarization shift from a pro-inflammatory to a regulatoryphenotype, the following experiments were carried out. Peritonealmacrophages were harvested from C57BL/6 mice by injectingintra-peritoneally 1 ml of 3% Brewer thioglycolate medium(Sigma-Aldrich, St Louis, Mo., USA). Four days later, mice weresacrificed and peritoneal cells were collected from the abdominal cavityby washing with 5 ml PBS. Cells (1.4×106 cells/ml) were dispensed onto6-well plates (Corning Costar, Corning, N.Y. USA) and incubated at 37°C. in 5% CO2 for 75 min. Non-adherent cells were discarded and RPMI-1640(Gibco, Grand Island, N.Y., USA) containing 10% fetal calf serum(Biological Industries, Kibutz, Beit Haemek, Israel) was added. For M1pro-inflammatory differentiation studies 10 ng/ml (GM-CSF) and 10 ng/mlLipopolysaccharides (LPS) (PeprotTech, Rocky Hill, N.J., USA) were addedto the culture medium followed by incubation at 37° C. in 5% CO2 for 20hrs. Alternatively, for M2 regulatory differentiation 10 ng/ml XXXX(M-CSF) and 10 ng/ml IL-4 (PeprotTech) were added to media. sPIF wasadded to the medium together with the differentiation factors.

Flow cytometry differentiated macrophages analysis: To determine theeffect of sPIF on M1 to M2 differentiation after 20 hrs of culturedifferentiated macrophages by (IL4 and GMSCF) were harvested by usingTrypsin-EDTA solution (Biological Industries, Israel). Cells werestained at 4° C. For intracellular staining, cells were fixed in 1%paraformaldehyde (Electron Microcopy Sciences Hatfield, PA USA) and thenpermeabilized by saponing (number SIGMA-Aldrich, St. Louis, Mo. USA).The following antibodies were used: anti-mouse CD11b APC(SouthernBiotech, Birmingham, Ala., USA), anti-mouse F4/80 Pacific Blue(BioLegend, San Diego, Calif., USA), anti-mouse CD206 FITC (AbD Serotec,Raleigh, N.C., USA) anti-mouse CD16/32 PE and anti-mouse CD23 eFluor 660(eBioscience). Flow cytometry was performed using the MACSQuant®analyzer (Miltenyi Biotech, San Diego, Calif., USA).

Example 1

PIF's protection against lethal radiation has been demonstrated (FIG.1A). Mice (C57BL/6, n=36) were treated with low-dose PIF (0.75 mg/kg) orhigh-dose PIF (1.25 mg/kg) 2×/day for 14 days starting 2 hours afterlethal 8Gy irradiation. Such an exposure led to 100% survival 2 weeksafter stopping therapy. In contrast, control mice (n=14 males; 7females) that received radiation and (PBS, vehicle), but no PIFtreatment, developed ARS and died by day 23 (0% survival). FIG. 1B showsthat the global WBC count was preserved at day 0 compared with day 9 inboth PIF-treated groups as compared with the control group, which by day12 already had a very low global WBC count. FIG. 1C shows data fromfemale mice (n=9, similar results in males) that were treated with PIF0.75 mg/kg 2×/day (low, high dose: 0.75 mg/kg) for 14 days starting 2hours after 8Gy radiation exposure. The top panel shows the immunephenotype measured at 0-19 days. The second panel shows the same immuneprofile expressed as % of total. The third panel shows detailed redblood cell indices (blood count, hemoglobin, hematocrit, and volume ofRBCs. The bottom panel has the platelet counts and volume. FIGS. 1D and1E show that in the PIF-treated groups, immune phenotype, RBC, andplatelet count were preserved when measured until day 29—two weeks afterstopping PIF administration.

Example 2

sPIF effect of irradiation on C57BL/6 mice survival: dose-findingexperiments: The C57BL/6 mice used in this study are a relativelyradio-resistant strain. First, the radiation intensity required for ourexperiments by evaluating the survival curve in mice exposed to variousdoses of total body irradiation (6-10 Gry) was determined (FIG. 2). Allmice (n=10) survived 30 days after exposure to 6 Gry while all mice(n=10) died within 30 days after exposure to 10 Gry. B. showed that PIFimproves platelet count following 7Gy exposure. Therefore, 6 Gry wasdefined as sub-lethal dose to determine sPIF's effect on the hematologicprofile, cytokine expression and gene expression and 10 Gry as lethaldose for BM transplantation studies. In addition, since PIF was testedfor the first time in an ARS setting, the following studies were carriedout by using sPIF as sole therapy and without the traditional use ofantibiotics which has an important role in infection preventionpost-radiation. As such, specific mechanisms involved in sPIF actioncould be clearly dissected.

Example 3

sPIF promotes hematologic recovery after sub-lethal irradiation forlong-term post-therapy: The consequence of sub-lethal irradiation islong-term hematopoietic suppression associated with a slow recovery.sPIF's ability to restore WBC profile was determined (FIG. 3A). EarlysPIF administration (1 mg/kg/day for 2 weeks) 24 hrs after 6Gryirradiation led to rapid recovery and significantly improvedreconstitution of total circulating WBC as compared with PBS control(FIG. 3B). This was already evident at 2 weeks after irradiation whenWBC have reached a mean value of 600 cells/μl in the sPIF treated groupas compared with 250 cells/μl in PBS treated group. Such a level of 600WBC/μl indicates that these mice were less immuno-compromised already atthis time point. Notably, the protective effect of sPIF expanded beyondthe 2-week treatment with significant differences observed also at 4weeks after irradiation 2 weeks post-therapy at the time of sacrifice(FIG. 3C). Moreover, 2 weeks post-administration, sPIF increased thelymphocyte count while reducing circulating granulocytes percentageclosely to levels observed in normal untreated group (control) (FIG.3D). In contrast, in the sham PBS-treated group, lymphocyte count waslower and granulocyte count increased significantly as compared with thesPIF-treated group. The reduction in neutrophil count—an inflammatoryresponse associated with the increase in lymphocytes found in anadaptive immune response—reflects a beneficial restored immune profileto that seen in normal mice. This is especially relevant since long-termthe lymphocyte population is affected following radiation leading tohigh vulnerability to infection. These results demonstrate sPIFeffectiveness in restoring systemic immune cells profile post-sub-lethalradiation long-term.

Example 4

Short-term sPIF reduces systemic pro-inflammatory cytokines level postsub-lethal irradiation: A systemic inflammatory response rapidly followsionizing radiation. Therefore, the effect of sPIF starting at 24 hrspost-6 or 7Gy irradiation and administered for only 3 days was examined.(FIG. 4A) sPIF led to a significant reduction in prime pro-inflammatorycytokines IL-2 and IL1α circulating levels as compared with PBS. (FIGS.4B and 4C). In addition, when compared with levels in normal mice nodifferences in both cytokine levels were noted. The effect on othercytokines was not significant. These results imply that the sPIF-inducedreduction in the systemic inflammatory milieu early on post-radiationplays an important role in the protection against ARS development.

Example 5

sPIF local and systemic protection: promotes colon crypts recoverypost-sub-lethal radiation: ARS rapidly leads to GI inflammation andinjury and sPIF improves GI tract ulcers and liver inflammation in aharsh murine GVHD model post-lethal irradiation long-term. The aboveexperiments substantiated both in vivo and in vitro that sPIF isprotective in reducing systemic inflammatory mediators leading tolong-term recovery. However, whether PIF's effect is also local, targetsan organ the colon that has a high cellular turn-over, and which isfrequently affected by ARS is not known. Short-term sPIF injections for2 or 3 days post-sub-lethal radiation (6Gry) starting treatment at 24 or48 hrs post-radiation protects against colon inflammation, asillustrated in FIG. 5A. The model used which shows that sPIF reducedsignificantly colon inflammation restoring colon crypts morphology(FIGS. 5B and 5C). Remarkably the protective effect was also observedwhen the sPIF treatment has started only at 48 hrs post-irradiation. D.Shows that PIF crypt recovery is similar to that of normal mice. E. PIFreduces iNOS-oxidative stress gene while promoting B7H1 protective genein the colon (RTPCR). The effect was noted when PIF treatment started at48 h hours post-therapy. This supports the view that sPIF exerts anintegrated both systemic (hematopoietic, cytokines) and localprotection.

Example 6

sPIF promotes colon recovery by reducing oxidative stress, modulatingmitochondrial function, immune response. The above data indicated thatsPIF has a dual local and systemic effect both reducing the damagingeffect of radiation restoring colon architecture while reducing thesystemic pro-inflammatory immune response enabling long-termhematopoiesis. Therefore using both RT PCR (FIGS. 5D and 5E) effect onNOS2, and B7H1 and global genome analysis (FIG. 10) we focused onexamining principally two intercalating items local protection and colonfunction. sPIF treatment (compared to sub-lethal) resulted insignificant modulation of several pathways (FIG. 10). Interestinglythese genes mainly cluster in three groups which can globally areinvolved in immune regulation and apoptotic signaling, intracellularenergy transfer, and Protein-RNA interactions required for effectivemetabolic function. The first two are related to protective effect whilethe latter is related to repair mechanisms.

Table 5 below details a cluster analysis of the global colon genome. Theanalysis compared sPIF vs PBS and normal mice. Several pathways wereaffected. Table 4 above shows individual pathways identified and theirranks. A cluster analysis of major pathways' mitochondrial function,response to stress and protein-RNA interactions was performed.(“Fluor”=Fluorescence Measurement). The gene names correspond to thegenes identified in the Illumina MouseRef-8 array Instrutions booklet(BD-202-0202, Illumina, USA), which is herein incorporated by referencein its entirety. In brief, following sub-lethal radiation (6Gry) at 24hrs sPIF injection twice daily for 72 hrs were administered. The effectof sPIF following radiation was compared with PBS treated control.Normal mice without irradation served as an additional control.Following sacrifice 30 mg of colon tissue (N=7 for PIF and PBS and N=3for normal mice) was excised and homogenized in a Fastprep 120 tissuehomogenizer (30 s at 4.0 m/sec) in cell lysis buffer (Qiagen,Hombrechtikon, Switzerland). Total RNAs were extracted from cells usingPureLink RNA Mini Kit (Ambion, catalog number 12183018A). Total RNA (250ng) was amplified into cRNA using TotalPrep RNA amplification kit(AMIL1791, Ambion) following manufacture's instruction. Afteramplification, 1.5 μg of cRNA was mixed with the hybridization controlsand it was hybridized to MouseRef-8 array (BD-202-0202, Illumina, USA).The array was hybridized for 16 hrs in a hybridization oven with arocking platform at 58° C. The array chip then went through a series ofwashes before it was stained with streptavidin-Cy3. After the staining,it went through a final wash and drying. The array was scanned using theIllumina HiScan Scanner.

TABLE 5 Normalized Normalized increase in decrease in expressionexpression level as level as Raw Ratio of compared Raw Ratio of comparedto Fluor. increase in to absence Fluor. decrease in absence of Gene nameValue expression of PIF Gene name Value expression PIF Cfd 11537 0.8581.57E−001 9.67E−001 Fabp6 16204 −3.060 3.83E−002 Olfm4 380924 0.7991.00E−001 9.67E−001 LOC100046120 NA −1.054 3.58E−002 Lyz1 17110 0.7333.20E−001 9.67E−001 Tmem117 320709 −0.788 2.03E−001 Scd1 20249 0.7301.21E−001 9.67E−001 Serpina1b 20701 −0.721 1.35E−001 Rnase1 19752 0.7158.96E−002 9.67E−001 Prss7 19146 −0.681 2.68E−001 Adh1 11522 0.7143.37E−001 9.67E−001 Fam151a 230579 −0.627 1.58E−001 Reg1 19692 0.6796.32E−001 9.75E−001 Pcsk9 100102 −0.610 5.22E−002 Cyp3a11 13112 0.6542.30E−001 9.67E−001 Cfl2 12632 −0.587 3.51E−001 Casp6 12368 0.6471.20E−001 9.67E−001 Mfge8 17304 −0.579 4.13E−002 Ccl5 20304 0.6478.73E−004 9.67E−001 Ccl21a 18829 −0.576 1.35E−001 Acaa2 52538 0.6181.39E−001 9.67E−001 Slc5a6 330064 −0.557 7.03E−002 Rps3 27050 0.6102.19E−002 9.67E−001 Xpnpep2 170745 −0.555 1.17E−001 Khk 16548 0.6004.04E−002 9.67E−001 Ddah1 69219 −0.554 2.67E−001 Mbl2 17195 0.5952.21E−002 9.67E−001 Cxcl13 55985 −0.535 4.91E−002 Atp5f1 11950 0.5861.52E−001 9.67E−001 Mfge8 17304 −0.506 9.26E−002 Scye1 13722 0.5741.82E−002 9.67E−001 Gm766 330440 −0.502 3.75E−001 Sep15 93684 0.5511.79E−001 9.67E−001 Serpina1b 20701 −0.497 1.07E−001 Ppp1ca 19045 0.5492.28E−001 9.67E−001 Osta 106407 −0.491 5.77E−002 Cox7a2l 20463 0.5395.23E−002 9.67E−001 Panx1 55991 −0.490 4.65E−003 Clca6 99663 0.5271.26E−002 9.67E−001 Zmiz1 328365 −0.488 2.21E−002 Apoa4 11808 0.5231.50E−001 9.67E−001 Aim1 11630 −0.480 1.18E−002 Calm2 12314 0.5184.17E−002 9.67E−001 Serpina1d 20703 −0.460 6.93E−002 Cyp2c65 72303 0.5161.44E−001 9.67E−001 Apaf1 11783 −0.459 1.51E−002 Hspa1a 193740 0.5101.21E−001 9.67E−001 Parp14 547253 −0.442 6.88E−002 Suclg1 56451 0.5092.09E−001 9.67E−001 Naaladl1 381204 −0.438 3.97E−001 Gsta2 14858 0.5072.97E−001 9.67E−001 Ccnd1 12443 −0.431 1.52E−001 Adipoq 11450 0.5069.44E−002 9.67E−001 Iap NA −0.428 1.37E−001 Cyp3a25 56388 0.5021.69E−001 9.67E−001 BC040758 268663 −0.412 1.02E−001 H2afz 51788 0.4871.40E−001 9.67E−001 Uba1 22201 −0.412 5.03E−002 Arg2 11847 0.4831.73E−001 9.67E−001 Anpep 16790 −0.406 6.74E−002 Tubb2b 73710 0.4807.79E−003 9.67E−001 LOC100040592 NA −0.406 4.13E−002 H3f3a 15078 0.4801.36E−001 9.67E−001 Coro1c 23790 −0.406 3.80E−002 Npm1 18148 0.4778.07E−002 9.67E−001 Pyy 217212 −0.405 4.02E−001 Rpl7a 27176 0.4731.62E−001 9.67E−001 Cd74 16149 −0.404 2.90E−001 Hspa8 15481 0.4723.60E−001 9.67E−001 Dag1 13138 −0.403 1.49E−002 Rnaset2 68195 0.4721.13E−001 9.67E−001 Mep1a 17287 −0.403 3.37E−002 EG433923 433923 0.4671.63E−001 9.67E−001 LOC100041504 1E+08 −0.403 1.20E−001 Acta2 114750.463 1.38E−001 9.67E−001 Gata5 14464 −0.403 1.01E−002 Cyp2b10 130880.462 3.82E−001 9.67E−001 Igsf3 78908 −0.399 8.49E−002 Cldn4 12740 0.4618.16E−003 9.67E−001 Sqle 20775 −0.397 2.23E−001 Cyb5 109672 0.4511.47E−002 9.67E−001 Myadm 50918 −0.390 5.39E−002 Cml4 68396 0.4505.36E−002 9.67E−001 Stat3 20848 −0.385 6.65E−002 Tsc22d1 21807 0.4368.28E−002 9.67E−001 Sec16a 227648 −0.385 3.59E−002 Pnliprp2 18947 0.4351.92E−001 9.67E−001 Nr1h4 20186 −0.383 1.21E−001 Lgsn 14661 0.4347.15E−002 9.67E−001 Lamb3 16780 −0.382 1.68E−002 Cyp2b23 243881 0.4307.79E−002 9.67E−001 Elf3 13710 −0.381 7.89E−002 Ndufc2 68197 0.4301.11E−001 9.67E−001 Mfi2 30060 −0.381 4.70E−002 Immp21 93757 0.4281.12E−001 9.67E−001 Npc1l1 237636 −0.379 1.81E−001 Sphk1 20698 0.4274.24E−002 9.67E−001 Brd4 57261 −0.376 1.08E−001 Ppa2 74776 0.4251.94E−003 9.67E−001 Iqgap2 544963 −0.375 2.07E−002 Sumo2 170930 0.4251.64E−001 9.67E−001 Dgka 13139 −0.373 4.27E−002 Rnu6 19862 0.4191.74E−001 9.67E−001 Dync1h1 13424 −0.371 1.47E−001 Dnajc19 67713 0.4181.13E−001 9.67E−001 Nucb1 18220 −0.371 1.47E−002 Mrp153 68499 0.4151.31E−001 9.67E−001 Reep3 28193 −0.371 4.81E−002 Tmem33 67878 0.4132.35E−001 9.67E−001 Hsd17b4 15488 −0.370 1.46E−002 Dnase1 13419 0.4095.11E−002 9.67E−001 Pml 18854 −0.368 8.76E−003 Npm3-ps1 108176 0.4073.07E−002 9.67E−001 Irf1 16362 −0.364 3.73E−002 Cbr1 12408 0.4051.88E−001 9.67E−001 Ogdh 18293 −0.363 1.78E−001 Ifi27 76933 0.4044.78E−002 9.67E−001 Gdpd1 66569 −0.361 2.61E−001 Gsto1 14873 0.4046.86E−002 9.67E−001 Midn 59090 −0.360 1.40E−001 Alg5 66248 0.4018.97E−002 9.67E−001 Xlr4a 434794 −0.359 3.30E−002 Tac1 21333 0.4013.26E−002 9.67E−001 Psap 19156 −0.358 6.97E−002 Thap4 67026 0.4007.65E−003 9.67E−001 Dag1 13138 −0.354 7.77E−003 Sdcbp 53378 0.3982.16E−001 9.67E−001 Hgs 15239 −0.354 8.72E−002 Hist1h1c 50708 0.3961.58E−003 9.67E−001 H2-Eb1 14969 −0.353 2.47E−001 Lgals6 16857 0.3934.66E−002 9.67E−001 Slc44a4 70129 −0.351 1.50E−001 Rps2 16898 0.3921.35E−001 9.67E−001 Gp1bb 14724 −0.350 2.10E−001 Fbp1 14121 0.3906.83E−002 9.67E−001 Rrbp1 81910 −0.350 6.50E−002 Rpl23 65019 0.3843.29E−002 9.67E−001 Ttyh3 78339 −0.350 6.93E−002 Gsta3 14859 0.3847.46E−002 9.67E−001 Ahnak 66395 −0.350 1.59E−001 Mbnl2 105559 0.3832.89E−003 9.67E−001 Bcl3 12051 −0.345 1.32E−002 Nme2 18103 0.3774.00E−002 9.67E−001 H2-DMa 14998 −0.344 2.82E−001 EG434858 434858 0.3762.46E−001 9.67E−001 Ceacam20 71601 −0.344 1.70E−001 Eef2 13629 0.3753.22E−001 9.67E−001 Mfsd7c 217721 −0.342 2.23E−001 Hint1 15254 0.3741.38E−001 9.67E−001 Sema4b 20352 −0.342 9.11E−002 Chpt1 212862 0.3731.01E−001 9.67E−001 Preb 50907 −0.341 8.95E−003 Naca 17938 0.3721.95E−001 9.67E−001 Smap2 69780 −0.340 2.16E−002 Hist1h2bf 319180 0.3701.09E−001 9.67E−001 Sema4a 20351 −0.339 6.92E−002 Gpx4 625249 0.3709.91E−002 9.67E−001 Cyp2s1 74134 −0.339 1.28E−001 Bat5 193742 0.3691.96E−002 9.67E−001 Kcnk6 52150 −0.338 1.65E−001 Gsdmdc1 69146 0.3681.51E−001 9.67E−001 Plec1 18810 −0.338 2.68E−001 Hist1h2bh 319182 0.3671.22E−001 9.67E−001 Mall 228576 −0.336 1.64E−001 Hnrnpa2b1 53379 0.3663.04E−001 9.67E−001 Abcd3 19299 −0.335 3.94E−002 Cyp2d26 76279 0.3622.72E−001 9.67E−001 Gapvd1 66691 −0.335 2.01E−002 Pigp 56176 0.3621.54E−001 9.67E−001 Gbf1 107338 −0.333 8.55E−002 Hpgd 15446 0.3606.36E−002 9.67E−001 Ext1 14042 −0.333 1.30E−002 Ndufs3 68349 0.3597.02E−002 9.67E−001 Ggt1 14598 −0.332 1.70E−002 Lypla1 18777 0.3593.18E−001 9.67E−001 Gpt1 76282 −0.332 8.65E−002 Slc2a2 20526 0.3581.44E−001 9.67E−001 Tcf7l2 21416 −0.332 6.88E−002 Ada 11486 0.3577.39E−002 9.67E−001 Tmc5 74424 −0.330 5.09E−002 Ndufa12 66414 0.3561.09E−001 9.67E−001 Lmtk2 231876 −0.329 3.25E−002 Gm1123 382097 0.3563.79E−001 9.67E−001 Acta1 11459 −0.328 8.55E−002 Fos 14281 0.3561.46E−002 9.67E−001 Slc35c1 228368 −0.327 7.77E−002 Sumo1 22218 0.3562.03E−001 9.67E−001 Ece1 230857 −0.325 9.16E−002 Tbrg1 21376 0.3552.44E−001 9.67E−001 Igf2 16002 −0.324 4.88E−001 Plekhf1 72287 0.3541.79E−001 9.67E−001 Camk2b 12323 −0.321 4.02E−002 Rps7 20115 0.3542.98E−001 9.67E−001 H2-DMb1 14999 −0.321 2.94E−001 Tbca 21371 0.3533.99E−002 9.67E−001 Zfp710 209225 −0.320 4.33E−003 Ufsp2 192169 0.3522.35E−001 9.67E−001 Neu1 18010 −0.320 2.52E−001 Prdx5 54683 0.3504.48E−003 9.67E−001 Ostb 330962 −0.319 2.55E−001 Ndufa6 67130 0.3485.79E−002 9.67E−001 Purb 19291 −0.317 1.13E−001 Mtch2 56428 0.3483.69E−001 9.67E−001 Gns 75612 −0.317 6.64E−002 Arrdc4 66412 0.3473.73E−002 9.67E−001 Rnasen 14000 −0.314 1.23E−002 Eif5 217869 0.3461.89E−001 9.67E−001 B230339M05Rik 228850 −0.313 4.26E−002 Eif3s4 533560.346 1.51E−001 9.67E−001 Slc9a1 20544 −0.312 2.61E−002 Fam109a 2317170.345 1.75E−003 9.67E−001 Tns4 217169 −0.312 4.28E−002 Arl6ip5 651060.345 6.55E−002 9.67E−001 Nos2 18126 −0.310 2.50E−001 Apoc2 NA 0.3442.07E−001 9.67E−001 Camta2 216874 −0.309 2.55E−001 Tspan3 56434 0.3432.79E−001 9.67E−001 Gpx2 14776 −0.307 6.23E−002 Ccl9 20308 0.3421.71E−001 9.67E−001 Gna13 14674 −0.307 3.80E−002 Psma7 26444 0.3421.65E−001 9.67E−001 Vcp 269523 −0.306 1.81E−001 Use1 67023 0.3421.15E−001 9.67E−001 Hipk2 15258 −0.305 1.39E−001 Sdhb 67680 0.3411.38E−001 9.67E−001 Xpnpep1 170750 −0.305 1.34E−001 Cd59a 12509 0.3412.97E−002 9.67E−001 Aff1 17355 −0.305 8.78E−002 Ak2 11637 0.3402.08E−001 9.67E−001 Adcy8 11514 −0.303 1.99E−001 Tmem85 68032 0.3406.90E−002 9.67E−001 LOC100048299 NA −0.302 9.56E−002 C1qbp 12261 0.3401.77E−001 9.67E−001 Abcf1 224742 −0.302 8.60E−002 Prdx4 53381 0.3381.84E−001 9.67E−001 Agpat4 68262 −0.299 2.01E−001 Hebp1 15199 0.3373.33E−004 9.67E−001 Ctdsp2 52468 −0.297 7.91E−002 Ap1s1 11769 0.3365.98E−002 9.67E−001 C4b 12268 −0.296 1.06E−001 Nudt19 110959 0.3352.44E−002 9.67E−001 Lrrc1 214345 −0.296 1.74E−002 Rdh7 54150 0.3354.26E−001 9.67E−001 NA −0.294 6.10E−002 9.67E−001 Adk 11534 0.3331.24E−001 9.67E−001 Chst8 68947 −0.294 2.30E−001 Rpl13a 22121 0.3331.18E−001 9.67E−001 Copz1 56447 −0.294 9.89E−002 Ppp2r5c 26931 0.3321.67E−001 9.67E−001 Clec2d 93694 −0.293 2.88E−001 Mbnl1 56758 0.3313.47E−002 9.67E−001 Dfna5h 54722 −0.293 4.73E−001 Rheb 19744 0.3317.93E−002 9.67E−001 Kif1b 16561 −0.293 6.76E−002 Mrpl9 78523 0.3302.14E−001 9.67E−001 Sec63 140740 −0.293 2.57E−002 Eif5a 276770 0.3303.72E−001 9.67E−001 Fntb 110606 −0.293 1.53E−001 Sar1b 66397 0.3292.56E−001 9.67E−001 Capg 12332 −0.293 1.04E−001 Rdh7 54150 0.3264.55E−001 9.67E−001 Sidt2 214597 −0.293 2.15E−001 Indo 15930 0.3251.40E−001 9.67E−001 Mboat1 218121 −0.292 2.99E−001 Oas1g 23960 0.3232.57E−001 9.67E−001 AI427809 381524 −0.292 1.20E−001 Gde1 56209 0.3231.81E−001 9.67E−001 Pcyt1a 13026 −0.291 1.69E−002 Atp5c1 11949 0.3208.74E−002 9.67E−001 Atp6ap1 54411 −0.290 3.13E−002 Idh3g 15929 0.3203.15E−001 9.67E−001 AI451617 209387 −0.289 6.12E−002 Atp5l 27425 0.3201.10E−001 9.67E−001 Atp6v0a1 11975 −0.288 2.29E−001 H2-Q2 15013 0.3191.28E−001 9.67E−001 Slco2a1 24059 −0.288 1.22E−001 Sepp1 20363 0.3191.47E−001 9.67E−001 Batf2 74481 −0.288 1.75E−002 Dcn 13179 0.3171.86E−001 9.67E−001 Dusp6 67603 −0.287 2.50E−001 Slc28a2 269346 0.3166.25E−002 9.67E−001 LOC100044566 NA −0.286 7.52E−002 Npm3 18150 0.3156.24E−003 9.67E−001 Papss2 23972 −0.285 3.48E−001 Timm8b 30057 0.3151.54E−001 9.67E−001 Gpd2 14571 −0.285 9.42E−002 S100a1 20193 0.3141.53E−001 9.67E−001 Mapk6 50772 −0.285 8.07E−002 Ddit4 74747 0.3131.42E−001 9.67E−001 Entpd5 12499 −0.284 2.76E−002 Tmem49 75909 0.3133.81E−001 9.67E−001 Sbf1 77980 −0.282 1.63E−002 Ntan1 18203 0.3126.37E−002 9.67E−001 Dlst 78920 −0.282 1.09E−001 Cfi 12630 0.3121.27E−001 9.67E−001 Lasp1 16796 −0.281 1.02E−001 Mocs2 17434 0.3121.11E−001 9.67E−001 Krt20 66809 −0.281 3.82E−002 Gsta1 14857 0.3114.90E−001 9.67E−001 Ncor1 20185 −0.281 1.62E−001 Rnase4 58809 0.3101.90E−001 9.67E−001 Serpina3n 20716 −0.279 1.40E−001 Tmem77 67171 0.3102.32E−001 9.67E−001 Gjb3 14620 −0.278 1.23E−001 Usp18 24110 0.3092.43E−001 9.67E−001 Srr 27364 −0.278 2.91E−002 Atp6v0e 11974 0.3081.71E−001 9.67E−001 Vdr 22337 −0.277 1.62E−001 Rplp0 11837 0.3081.58E−002 9.67E−001 Sppl3 74585 −0.276 2.20E−001 Vps29 56433 0.3071.59E−001 9.67E−001 Sgk1 20393 −0.276 2.69E−001 Rdh14 105014 0.3061.27E−001 9.67E−001 Rnf185 193670 −0.276 1.08E−001 Mep1b 17288 0.3043.36E−001 9.67E−001 Rfx1 19724 −0.275 7.80E−002 Mrps33 14548 0.3032.22E−001 9.67E−001 Epb4.1l1 13821 −0.275 9.00E−002 Nipsnap3a 665360.303 1.30E−001 9.67E−001 Cpxm1 56264 −0.274 9.20E−002 Dnajc15 661480.302 2.42E−001 9.67E−001 Sgk2 27219 −0.274 1.51E−001 Hint3 66847 0.3011.51E−001 9.67E−001 Specc1l 74392 −0.274 1.51E−001 Eif3g 53356 0.3002.20E−001 9.67E−001 Tmem50a 71817 −0.274 6.52E−004 Rps27a 78294 0.3003.28E−001 9.67E−001 Utx 22289 −0.273 1.87E−001 Akr1c12 622402 0.3004.27E−001 9.67E−001 Fam102a 98952 −0.272 1.79E−001 Spcs1 69019 0.2992.37E−001 9.67E−001 Kctd5 69259 −0.272 6.67E−002 Hmgn2 15331 0.2991.29E−001 9.67E−001 Rtn3 20168 −0.272 1.00E−002 Ccl11 20292 0.2992.69E−001 9.67E−001 Grit 330914 −0.271 2.24E−001 Tyms 22171 0.2982.79E−001 9.67E−001 Csnk1d 104318 −0.270 2.78E−002 Bpnt1 23827 0.2978.58E−002 9.67E−001 Flt1 14254 −0.270 1.16E−001

Example 7

sPIF prevents colon inflammation, Down-regulates nitric oxide (NOS2) andup-regulates HSPs and B7H1 expression to enable metabolic function: Thecolon histology (FIG. 5B) indicated that sPIF protects againstinflammation by restoring the colon crypts. sPIF plays a major role inprotecting against oxidative stress and nitric oxide (NO) formationthrough eNOS (NOS2) pathways in both liver and in macrophages. sPIFdown-regulated the NOS2 gene expression in the colon as well (FIG. 5E).By promoting NOSIP gene NO production decreased as NOS1 and NOS3 aretranslocated to the actin cytoskeleton attenuating theses enzymes'activity. The increased PTS (6-Pyruvoyltetrahydropterin Synthase), DDAH1and SPR and decreased WASL (Wiscott-Aldrich syndrome) genes furtherlimits NOS2 activity. In addition, NDOFA12 and 6 regulatorymitochondrial membrane respiratory chain NADH dehydrogenases noncatalytic subunits expression decreased as well. Beyond the reducedoxidative stress, sPIF also prevents protein misfolding by increasingHSPa1a and HSPA8 genes also known to be targeted by sPIF and is alsoregulated in vivo. The local colon innate immune system as shown by B7H1expression is independent of the systemic immunity and B7H1 and wasreported as a prime protector against colon inflammation. Remarkably,sPIF promoted B7H1 gene expression as compared with control (FIG. 5E).The increase in B7H1 was higher than in sham, indicating a potentprotective response. Significant protection was also noted when sPIFtherapy was started only 48 hours post-radiation. The highest expressedgene is CFD a complement D factor which protects against infection theconsequence of colon damage. This is coupled by the effect of OLFM4which has an additive antiapoptotic effect. To favor local metabolismthe upregulation of SCD1 and CYP3A11 and ATP5fi provide the energyrequired for such an important task. Among leading genes upregulated wasACAA2, which is involved in fatty acid and selective amino acidsmetabolism and KHK which is involved in fructose metabolism amongothers. Slc5a6 expression, which is involved in solute transport, wasalso upregulated. This data indicates that sPIF-induced colon protectionis associated with enhanced metabolism promoting genes expression.

Example 8

sPIF enhances hematopoietic recovery post-lethal irradiation followed bysemi-allogeneic BMT: For patients exposed to high (lethal) levels ofradiation, hematopoietic stem cell transplantation is routinelyconsidered. Since a donor must be available in a short notice,haplo-identical allogeneic transplantation might be the only option. Itwas therefore decided to evaluate the effect of sPIF on hematologicrecovery post-lethal total body irradiation followed by semi-allogeneicBMT, which resembles parent-to-child transplantation. In order to mimica clinical setting where a graft-versus-host reaction would begenerated, F1 (C57BL/6×Balb/c) mice were exposed to 10 Gry total bodyirradiation followed by intravenous administration of C57BL/6 BM cellsthe next day. (FIG. 6A) sPIF (1 mg/kg/day) or PBS was administeredcontinuously (0.25 ml/h) starting at 24 hrs post-irradiation, for twoweeks, using Alzet osmotic pumps. The clinical condition and hematologicrecovery was followed up to 4 weeks post-irradiation. This experimentaimed in addition to demonstrate whether sPIF prevents a reduction inWBC and/or it is also effective in BM repopulation restoringhematopoiesis. Three weeks post-irradiation following BMT, sPIFsignificantly increased the recovery of total WBC in the peripheralblood as compared to PBS treated group (FIG. 6B). In addition, sPIFtreatment improved the lymphocyte/granulocyte ratio as compared withPBS-treated control mice (FIG. 6C). This WBC ratio was similar to thatfound following sub-lethal irradiation. To further evaluate the effectof sPIF on hematologic recovery, histological examination of the femurbone was also performed. Significant difference in bone marrowcellularity was observed at 4 weeks post-irradiation and BMT at 2 weekspost-therapy. Representative histological images of the bone marrow fromfemur bone of normal, PBS and PIF treated mice are presented in FIGS.6D, 6E, and 6F, respectively. The fat cells number in the femur BMsections of sPIF treated group was significantly lower as compared toPBS control mice (FIG. 6G), indicating improved rehabilitation of the BMcells in the sPIF treated group. Remarkably, the number of fat cells inthe sPIF-treated group was similar to that observed in normal mice. Suchobservations indicate that post-lethal irradiation BMT coupled with PIFcan help rapidly restore both circulating as well the bone marrowreservoir

Example 9

Transplantation of sPIF-pre-treated allogeneic bone marrow enhanceshematologic reconstitution after lethal irradiation long term: sPIFregulates immune response. Therefore, whether sPIF has a directinfluence on transplanted bone marrow cells improving their engraftmentwas examined. Or alternatively sPIF leads to hematologic reconstitutiononly by exerting its immune-regulatory properties on the recipient bypromoting BMT engraftment. To address such critical question, donor BMcells (allogeneic) were pre-incubated with sPIF for 2 hrs in culture andthen the cells were washed off prior transplantation. Recipients werelethally irradiated with (10 Gry) and 24 hrs later transplanted with thepre-conditioned BM graft. No additional treatment was administered.(FIG. 7A)

Although the BM was incubated with sPIF for only 2 hrs prior totransplantation, the experimental group receiving sPIF-treated cellsshowed enhanced reconstitution of the total WBC count, and specificallylymphocyte count, three and four weeks after transplantation (FIGS. 7Band 7C). Whether this effect was exerted by improved bone marrowcellularity was further examined (FIGS. 7D, E, F). Data showed that sPIFled to restored BM by reducing fat cells presence in the femur. SincesPIF-based preconditioning was effective in vivo following BMT, thepossible mechanisms involved in this protection were further examined.The effect of sPIF pre-treatment on BM antigenicity was examined. sPIFpreincubated with MSCs for 2 hrs were added in culture to CFSE labelledsplenocytes activated by anti-CD3 antibody assessing effect onproliferation. (FIG. 7G) Flow cytometry data documented that suchprecondition has a significant effect reducing the number ofproliferating cells. FIG. 7F shows that PIF preconditioning of MSCs leadto their differentiation to B and T cells. FIG. 7H shows that PIFprevents weight loss when examined 5 days after transplantation.

This demonstrated that short-term exposure of sPIF for BM cells issufficient to lead to effective engraftment without requiring furthertherapy post-transplant long-term. This implies that short-term directeffect of sPIF in culture can translate into long-term pro-engraftmenteffects in vivo.

Example 10

sPIF alters macrophage differentiation: Given the importance of immuneresponse in ARS and immune modulatory effect of sPIF on monocytes,sPIF's effect on macrophages in vitro was tested. It was previouslyshown that PIF upregulates B7H1 in macrophages, reflecting immuneregulatory effects on T-cells proliferation. This upregulation wasfurther confirmed since in sPIF primed macrophages following co-culturewith activated T-cells has led to the reduced proliferation. The currentexperiments aimed to examine whether the protective effect of sPIF inboth sub-lethal and in lethal ARS models are due to a shift inmacrophage polarity thereby reducing the inflammatory response followingionizing radiation. Macrophages are key mediators of the immune responseand can differentiate into inflammatory (M1) and regulatory (M2)macrophages. Thus, we obtained peritoneal macrophages from C57BL/6 miceand differentiated them towards a M1 or M2 phenotype in the presence ofsPIF (FIGS. 8 and 9). Expectedly, sPIF significantly decreased iNOS(NOS2) and COX-2 genes expression (FIGS. 8A and 8B, respectively) in M1differentiating macrophages and enhanced the expression of arginase-1,which is a marker of M2 regulatory macrophages (FIG. 8C). Additionally,FACS analysis confirmed down-regulation of macrophage cell surface CD11band F4/80 expression to resemble an M2 macrophage phenotype (FIGS. 8Dand 8E). FIG. 9 shows that PIF shift CD16/32 and CD206 expression,further evidencing the shift to M2. Collectively, these results suggestthat sPIF may regulate immune response in irradiated mice by targetingM1/M2 macrophages.

PIF protects against lethally irradiated heart. I/1. Radiation-inducedheart disease (RIHD) is a concern during radiotherapy. Patho-mechanismsinvolved are progressive atherosclerosis of coronary arteries due toendothelial damage, and the diffuse injury of the myocardium due to theloss of small vessels and cardiomycytes replaced by fibrosis). In amurine model, the parallel study of the macro-vasculature-mediated anddiffuse radiation heart injuries with heart function studies(cardio-echo), the time-dependent evaluation of survival weight, skinhealing were evaluated Newborn rats 10-12/group were exposed to 50Gylethal radiation targeting the heart specifically. Subsequently sPIFcontinuous delivery was implanted using an Alzet pump. The pump releasedPIF for 2w 1 mg/kg/day. (The sketch of the protocol is described in FIG.13). Faster hair growth was noted on the site of irradiation and surgeryin the sPIF-treated animals than in the irradiated rats. The scarhealing was faster some days after the implantation of the Alzet pump inthe PIF-treated animals after 2 weeks long PIF-treatment. The dailyactivity of the PIF-treated animals was higher, they were much moreplayful then the other animals in the irradiated group.

Example 11

PIF core peptide sequence investigation: In silico docking of PIFwtreveal that chain A of the KCNAB1 (Kv1.3β) tetramer to be more likelyPIF target than the three other chains. The in silico mutagenesis ofdocked PIFwt: Kv1.3β interface also reveals M*RIKP*N importance(mutation in M1, I4, K5, P6 and N10 are strongest interface disruptors),but shows no similarity among the tetramer domains: a. FlexPepDockserver docking of PIF suggested that PIF is more stable when binding tochain A; b. BeatMusic server was used for in silico mutagenesis, inorder to predict which amino acid of the PIF sequence upon mutationwould yield a higher Energy PIF-Kv1.3β:A-D complex, and thus less stablestructure. Thus 2 putative mutants were conceived, mutant—1: P6->E6, andmutant—3: I4->G4; c. Table represent the mutants that were considered,of them 2 were synthesized based on their putative availability asoptions for ligand-receptor disruptors. Mutant 1 (P6->E6) is morespecific for chain D of the Kv1.3β, the form that also had high energyof binding and interface score. Mutant 3 (I4->G4) is predicted todisrupt the binding with chain A of the Kv1.3β. An example with moredetails of the in silico mutagenesis using several PIF targets alsosuggest that RIKP is the core sequence. The validation of the mutants isshown in FIG. 11 for another target of PIF—IDE (data on the DifferentialShift Assay is shown). The validation of mutants is also shown in FIG.12, which shows Differential Shift Assay data for Kv1.3β.

Flexible in silico docking (Algorithm #2: CABS Dock—this is a molecularmodelling approach where the PIF sequence is only provided, not a model,and the simulation of its molecular dynamics and flexibility are used toseek where the peptide would bind) models were compared to flexible insilico docking (Algorithm #1: FlexPepDock) by projecting PIF wt fromFlexPepDock over the FLAG-GG-PIF-HA models, obtained in CABS Dock.Remarkably, both algorithms (very distinct in nature) predict binding tothe same “pocket” in Kv1.3β with the consensus sequence RIKP shared inthe interface of both types of models, but with opposite directions. Thedata suggest double tagged PIF to be able to bind PIF wt targets in mostcases. IDE binding and PIFwt vs PlFmut-1/3 bound to Kv1.3β were studied.

Example 12

FIG. 14 showed that following 2w continuous and subsequent twice weeklyinjections sPIF improves the rats' weight as compared with control after19 weeks of the study. The effect was significant vs vehicle treatedrats. The growth curve was similar to non-irradiated rats.

FIG. 15 shows that post-lethal irradiation sPIF survival is preserveduntil the end of the study it is slightly lower than that in PBS treatedrats.

FIG. 16 described the cardiac indices that sPIF can affect followinglethal irradiation.

FIG. 17 shows that sPIF protects against lethal radiation affecting anumber of cardiac indices among them. AWTs-anterior wall thickness bothin systole and diastole. In addition it reduces PWT thickness duringdiastole, P<0.05.

FIG. 18 shows that sPIF significantly improved rats' body weight ascompared with PBS treated controls.

FIG. 19 shows that sPIF increase kidney weight as compared with PBScontrol. P<0.05

PIF injection protects against heterotropic cardiac graft rejection.Mice Balb/c were transplanted cardiac allograft to the abdominal cavityof the recipient (57B1/6) attaching the vessels to the abdominal aorta.Following transplant sPIF twice daily injections were carried outdetermining transplanted heart activity by determining cardiacpulsations using a stethoscope. The effect of sPIF was compared with PBStreated control. Data showed that sPIF delayed significantly rejection9+/−0.52 SE vs 7+/−0 (N=18 sPIF and N=5 control), P<0.002, (DF 17).

PIF promotes primary adrenal cell viability and cortisol secretion:basis for bioartificial adrenal. Congenital adrenal hyperplasia (CAH)due to deficiency of 21-hydroxylase is the most common genetic endocrinedisorder in humans, presenting with clinical symptoms of neuroendocrineperturbations, virilization and metabolic disease in later life.Patients may suffer from hypotensive crises, hypoglycemia, acne andinfertility.

Current treatment options with glucocorticoid substitution can reversethe symptoms only partially and exhibit the unpleasant side effects ofexcess glucocorticoid treatment. Despite of different treatmentalgorithms, the management of CAH remains a major therapeutic challengesometimes requiring drastic therapeutic measures such as bilateraladrenalectomy (Merke et al., 1999). Adrenal cell transplantation is afeasible therapeutic alternative for those patients. However, thisstrategy is critically limited by persistent lack of human donor organsand the requirement of chronic immune suppression.

One of the ways to solve those problems could be transplantation ofxenogeneic cells. Remarkable breakthrough in xenotransplantation isbased on application of microencapsulated in alginate xenogeneic cells.This method provides promising platform for cell therapy (Dolgin et al.,2014). Cell microencapsulation aims to protect the transplanted cellsfrom attack by the host immune system without immunosuppressive agents(Neufeld et al., 2013) and the recipient from immunization. There is afuture advantage that implantation could be accomplished by simpleinjection procedure rather by a surgical operation. PIF exposure toprimary bovine adrenal cells could be a suitable source for enablingeffective adrenal transplantation.

Experiments on primary bovine adrenocortical cells (BAC), isolated fromfour bovine adrenal glands. For viability, apoptosis and proliferationassay BAC were seeded in 96 well plates (1×10⁴ cells per well,sixplicate). For cortisol production assay cells were seeded in 24 wellplate (5×10⁴ cells per well, triplicate). PIF was dissolved in Salineand was used in concentration 0.1 μg/ml. One group of cells received PIFcontained medium right after cell isolation (PIF d0). Another group ofcells received PIF containing medium 24 hour post cell isolation (PIFd1). Control cells did not receive PIF. Viability was assessed using XTTCell Proliferation Assay (Roche) on day 3 after beginning of thetreatment. Proliferation was measured using BrdU Cell ProliferationAssay (Millipore) on day 3 after beginning of the treatment. Apoptosiswas assayed by determination of caspase 3/7 activity using Caspase-Glo3/7 Assay (Promega) on day 3 after beginning of the treatment. Forstimulated cortisol ACTH (Synacthen) in concentration 3 ng/mL was used.Cortisol in supernatant will be measured by EIA (IBL). Data in FIG. 20shows that PIF in culture increases cells viability while reducingapoptosis. Data in FIG. 21 shows that PIF promotes cortisol secretionsignificantly by these cells effect was most pronounced one day afteraddition to culture. This supportive data indicates that adrenal cellspre-conditioning could be valuable prior to transplant to the hosttherefore opens the possibility of developing a bio-artificial adrenal.

Experiments on rat insulinoma cells using PIF—basis for islettransplantation. PIF regulates global immune response both in vitro andin vivo [15-17]. PIF prevents diabetes development in NOD mice in bothadoptive transfer and spontaneously developing disease long term. PlFscrtested in parallel has no effect. The preserved islet architecture andinsulin expression is associated with increased pancreaticPDI/Thioredoxin and HSP proteins levels. This is coupled with systemicimmunity regulation reflected by changes in both TH1 and Th2 cytokinelevels. As recently shown (Barnea PLoS One 2014). PIF targets directlyPDI/T and HSPs through a shared binding site. Thus the protectionobserved against oxidative stress and protein misfolding critical fortransplant protection is mechanistically plausible. Having the abilityto transplant viable islets cells to patients with Type I diabetes wouldbe a major progress since until present such an approach is limited bothby the availability of donor cells as well after transplantation thereis a requirement for continuous immune suppression and despite theengraftment the transplanted cells fail to provide adequate insulin forthese patients on a long term basis.

PIF administration Ins-1 cells of passage 24 were used in theexperiments. For viability, apoptosis and proliferation assay cells wereseeded in 96 well plates (1×10⁴ cells per well, sixplicate). PIF wasused in three concentrations: 0.01 μg/ml; 0.1 μg/ml and 1 μg/ml. Medium,containing substrates, was changed every day. Viability was assessedusing XTT Cell Proliferation Assay (Roche). Proliferation was measuredusing BrdU Cell Proliferation Assay (Roche). Apoptosis was assayed bydetermination of caspase 3/7 activity using Caspase-Glo 3/7 Assay(Promega). Results were analysed by regression analysis for evaluationof the processes, occurring in the tested models. For evaluation of theeffects of PIF on cells Wilcoxon signed-rank test for related samplesand Spearman's rank correlation and Student's t-test were used. FIG. 22shows that PIF promotes insulinoma cells viability at 48 hours andproliferation at low 0.1 μg/ml concentration. The increase in apoptosisindicates elimination of cells of poor quality of frequent occurrence inculture conditions.

1. A method of treating and/or preventing acute radiation syndrome in asubject in need thereof following exposure to radiation comprisingadministering to the subject a therapeutically effective amount of a PIFpeptide, mimetics thereof, pharmaceutically acceptable salts thereof, orcombinations thereof.
 2. The method of claim 1, wherein the subject doesnot receive a bone marrow transplant.
 3. The method of claim 1, whereinthe therapeutically effective amount is from about 0.10milligrams/kilograms/day to about 10.00 milligrams/kilograms/day.
 4. Themethod of claim 1, wherein the therapeutically effective amount is fromabout 0.75 milligrams/kilograms/day to about 1.50milligrams/kilograms/day.
 5. The method of claim 1, wherein thetherapeutically effective amount is from about 0.75milligrams/kilograms/day to about 1.00 milligrams/kilograms/day.
 6. Themethod of claim 1, wherein the PIF peptide is administered within about24 hours after exposure to radiation.
 7. The method of claim 1, whereinthe PIF peptide is administered intermittently or continuously for atime period of from about 2 to about 14 days.
 8. The method of claim 1,wherein, if the PIF peptide is administered intermittently, the dosingregimen comprises about 1 dose per day or about 1 dose every two daysfor at least about twelve weeks.
 9. The method of claim 1, wherein thePIF peptide is administered intravenously, intramuscularly, orsubcutaneously.
 10. The method of claim 1, wherein the PIF peptide isadministered in an intermittent or continuous infusion.
 11. The methodof claim 1, wherein the acute radiation syndrome is caused by exposureto lethal or sub-lethal radiation.
 12. The method of claim 1, whereinthe acute radiation syndrome is caused by exposure to a radiation doseof about 100 rads to about 6000 rads.
 13. The method of claim 1, whereinthe acute radiation syndrome comprises delayed effects of acuteradiation exposure, including damage to any organ, tissue, or cell. 14.The method of claim 1, wherein the PIF peptide comprises one or acombination of an amino acid sequence having at least 86% sequencehomology with: SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, SEQ ID NO: 4,SEQ ID NO: 5, SEQ ID NO: 6, SEQ ID NO: 7, SEQ ID NO: 8, SEQ ID NO: 9,SEQ ID NO: 10, SEQ ID NO: 11, SEQ ID NO: 12, SEQ ID NO: 13, SEQ ID NO:14, SEQ ID NO: 15, SEQ ID NO: 16, SEQ ID NO: 17, SEQ ID NO: 18, SEQ IDNO: 19, SEQ ID NO: 20, SEQ ID NO: 21, SEQ ID NO: 22, SEQ ID NO: 23, SEQID NO: 24, SEQ ID NO: 25, SEQ ID NO: 26, SEQ ID NO: 27, SEQ ID NO: 28,SEQ ID NO: 29, mimetics thereof, and pharmaceutically acceptable saltsthereof. 15.-40. (canceled)
 41. A method of treating and/or preventingacute radiation syndrome following radiation exposure comprisingtransplanting one or a plurality of bone marrow cells into a subject inneed thereof, wherein the one or plurality of bone marrow cells ispre-exposed to a therapeutically effective amount of PIF peptide priorto transplantation, and the PIF peptide comprises or consists of anamino acid sequence having at least 86% sequence homology to an aminoacid sequence selected from: SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3,SEQ ID NO: 4, SEQ ID NO: 5, SEQ ID NO: 6, SEQ ID NO: 7, SEQ ID NO: 8,SEQ ID NO: 9, SEQ ID NO: 10, SEQ ID NO: 11, SEQ ID NO: 12, SEQ ID NO:13, SEQ ID NO: 14, SEQ ID NO: 15, SEQ ID NO: 16, SEQ ID NO: 17, SEQ IDNO: 18, SEQ ID NO: 19, SEQ ID NO: 20, SEQ ID NO: 21, SEQ ID NO: 22, SEQID NO: 23, SEQ ID NO: 24, SEQ ID NO: 25, SEQ ID NO: 26, SEQ ID NO: 27,SEQ ID NO: 28, SEQ ID NO: 29, mimetics thereof, and pharmaceuticallyacceptable salts thereof, and combinations thereof. 42.-44. (canceled)45. The method of claim 41, wherein the therapeutically effective amountof PIF peptide is from about 0.10 milligrams/kilograms/day to about10.00 milligrams/kilograms/day.
 46. The method of claim 41, wherein theone or a plurality of bone marrow cells is pre-exposed to aconcentration of PIF peptide for a time period and under conditionssufficient to pre-condition the bone marrow cells prior totransplantation and/or stimulate cortisol secretion. 47.-49. (canceled)50. A method of treating and/or preventing adult or juvenile type I ortype II diabetes comprising transplanting one or a plurality ofpancreatic islet cells into a subject in need thereof, wherein the isletcells are pre-exposed to a therapeutically effective amount of PIFpeptide prior to transplantation, and wherein the PIF peptide comprisesor consists of an amino acid sequence having at least 86% sequencehomology with: SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, SEQ ID NO: 4,SEQ ID NO: 5, SEQ ID NO: 6, SEQ ID NO: 7, SEQ ID NO: 8, SEQ ID NO: 9,SEQ ID NO: 10, SEQ ID NO: 11, SEQ ID NO: 12, SEQ ID NO: 13, SEQ ID NO:14, SEQ ID NO: 15, SEQ ID NO: 16, SEQ ID NO: 17, SEQ ID NO: 18, SEQ IDNO: 19, SEQ ID NO: 20, SEQ ID NO: 21, SEQ ID NO: 22, SEQ ID NO: 23, SEQID NO: 24, SEQ ID NO: 25, SEQ ID NO: 26, SEQ ID NO: 27, SEQ ID NO: 28,SEQ ID NO: 29, mimetics thereof, pharmaceutically acceptable saltsthereof, or combinations thereof.
 51. The method of claim 50, whereinthe subject has juvenile Type I diabetes.
 52. The method of claim 50,wherein the subject has adult Type I diabetes.
 53. (canceled)
 54. Themethod of claim 50, wherein the one or a plurality of pancreatic isletcells is pre-exposed to a concentration of PIF peptide for a time periodand under conditions sufficient to stimulate cortisol and/or insulinsecretion of the pancreatic islet cells prior to transplantation. 55.(canceled)
 56. The method of claim 50, wherein the one or plurality ofpancreatic islet cells is pre-exposed to a concentration of one or acombination of PIF peptides from about 10 nM to about 1000_ nM for about1 hour to about 4 hours.
 57. (canceled)
 58. A method of increasing thelikelihood of successful engraftment of a transplanted organ, tissue, orcells comprising transplanting an organ, tissue, or cell into a subjectin need thereof, wherein the organ, tissue, or cell is exposed to a PIFpeptide prior to transplantation, wherein the PIF peptide comprises orconsists of an amino acid sequence having at least 86% sequence homologyto an amino acid sequence chosen from: SEQ ID NO: 1, SEQ ID NO: 2, SEQID NO: 3, SEQ ID NO: 4, SEQ ID NO: 5, SEQ ID NO: 6, SEQ ID NO: 7, SEQ IDNO: 8, SEQ ID NO: 9, SEQ ID NO: 10, SEQ ID NO: 11, SEQ ID NO: 12, SEQ IDNO: 13, SEQ ID NO: 14, SEQ ID NO: 15, SEQ ID NO: 16, SEQ ID NO: 17, SEQID NO: 18, SEQ ID NO: 19, SEQ ID NO: 20, SEQ ID NO: 21, SEQ ID NO: 22,SEQ ID NO: 23, SEQ ID NO: 24, SEQ ID NO: 25, SEQ ID NO: 26, SEQ ID NO:27, SEQ ID NO: 28, SEQ ID NO: 29, mimetics thereof, and pharmaceuticallyacceptable salts thereof, and combinations thereof.
 59. The method ofclaim 58, wherein the transplantation is a bone marrow transplant,pancreatic islet cell transplant, adrenal cell transplant, or hearttransplant. 60.-63. (canceled)
 64. The method of claim 58, wherein theone or a plurality of organ, tissue, or cells is exposed to aconcentration of PIF peptide for a time period and under conditionssufficient to pre-condition the organ, tissue, or cells fortransplantation prior to transplantation. 65.-66. (canceled)
 67. Themethod of claim 58, wherein the one or plurality of pancreatic isletcells is pre-exposed to a concentration of one or a combination of PIFpeptides from about 10 nM to about 1000_ nM for about 1 hour to about 4hours.
 68. (canceled)
 69. The method of claim 58, wherein the organ,tissue, or cells are selected from one or a combination of organs chosenfrom: bone marrow, skin, adrenal gland, pancreas, heart, lung, kidney,spleen, and liver.
 70. A method of increasing the viability of an organ,tissue, or cell prior to its transplantation into a subject in need oftransplantation comprising treating the organ, tissue or cell with atherapeutically effective amount of PIF peptide prior totransplantation, wherein the PIF peptide comprises or consists of anamino acid sequence having at least 86% sequence homology to an aminoacid sequence chosen from: SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, SEQID NO: 4, SEQ ID NO: 5, SEQ ID NO: 6, SEQ ID NO: 7, SEQ ID NO: 8, SEQ IDNO: 9, SEQ ID NO: 10, SEQ ID NO: 11, SEQ ID NO: 12, SEQ ID NO: 13, SEQID NO: 14, SEQ ID NO: 15, SEQ ID NO: 16, SEQ ID NO: 17, SEQ ID NO: 18,SEQ ID NO: 19, SEQ ID NO: 20, SEQ ID NO: 21, SEQ ID NO: 22, SEQ ID NO:23, SEQ ID NO: 24, SEQ ID NO: 25, SEQ ID NO: 26, SEQ ID NO: 27, SEQ IDNO: 28, SEQ ID NO: 29, mimetics thereof, and pharmaceutically acceptablesalts thereof, and combinations thereof.
 71. A method of increasing thelikelihood of acceptance of a transplant of a donor organ, tissue, orcell into a subject, the method comprising exposing the organ, tissue orcell to one or more compositions comprising at least one PIF peptide ora mutant thereof or a pharmaceutically acceptable salt thereof prior totransplanting the organ, tissue, or cell into the subject. 72.-98.(canceled)